Difference between revisions of "Team:ZJU-China/Safety"
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<!-- <link href="assets/css/style.css" rel="stylesheet"> --> | <!-- <link href="assets/css/style.css" rel="stylesheet"> --> | ||
| − | <link rel="stylesheet" type="text/css" href="https://2020.igem.org/wiki/index.php?title=Template:ZJU-China/CSS/barcss/site&action=raw&ctype=text/css" /> | + | <link rel="stylesheet" type="text/css" href="https://2020.igem.org/wiki/index.php?title=Template:ZJU-China/CSS/barcss/site.css&action=raw&ctype=text/css" /> |
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<ul> | <ul> | ||
<li> | <li> | ||
| − | <a href="#Cloning" style="background-color: #f6b37f;" class="nav-active rounded"><span>Overview</span></a> | + | <a href="#Cloning" style="background-color: #f6b37f;" |
| + | class="nav-active rounded"><span>Overview</span></a> | ||
</li> | </li> | ||
<li> | <li> | ||
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</li> | </li> | ||
<li> | <li> | ||
| − | <a href="#services" style="background-color: #f19ec2;" class="rounded"><span>Insurance of Biosafety</span></a> | + | <a href="#services" style="background-color: #f19ec2;" class="rounded"><span>Insurance of |
| + | Biosafety</span></a> | ||
</li> | </li> | ||
<li> | <li> | ||
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</li> | </li> | ||
<li> | <li> | ||
| − | <a href="#respon" style="background-color:#d197f7;" class="rounded"><span>Always be Responsible</span></a> | + | <a href="#respon" style="background-color:#d197f7;" class="rounded"><span>Always be |
| + | Responsible</span></a> | ||
</li> | </li> | ||
<li> | <li> | ||
| − | <a href="# | + | <a href="#references" style="background-color: #cce198" class="rounded"><span>References</span></a> |
</li> | </li> | ||
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| − | <a class="nav-link" href="#">Team</a> | + | <div class="container"> |
| − | <ul class="dropdown-list"> | + | <a class="navbar-brand" href="https://2020.igem.org/Team:ZJU-China" style="left:-10%;margin-top:-18px;margin-bottom:-26px;margin-left:-18px;margin-right:-18px;"><img src="https://static.igem.org/mediawiki/2020/d/dd/T--ZJU-China--wiki_logo_new.png" class="img-fluid" width="150px" ></a> |
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| + | |||
| + | </li> | ||
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| + | <a class="nav-link" href="#">Team</a> | ||
| + | <ul class="dropdown-list" > | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Team">Members</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Attributions">Attributions</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Medals">Medals</a></li> | ||
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| + | |||
| + | </ul> | ||
| + | </li> | ||
| + | <li class="nav-item dropdown-box" style="background-color:cornflowerblue;;opacity: 0.9;"> | ||
| + | <a class="nav-link" href="#">Project</a> | ||
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| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Background">Background</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Design">Design</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Magnetosome">Magnetosome</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Experiments">Experiments</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Results">Results</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Future">Future</a></li> | ||
| + | </ul> | ||
| + | </li> | ||
| + | <li class="nav-item dropdown-box" style="background-color:dodgerblue;opacity: 0.9;"> | ||
| + | <a class="nav-link" href="#">Model</a> | ||
| + | <ul class="dropdown-list"> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Model">Model</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Engineering">Engineering</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Proof_Of_Concept">Proof of Concept</a></li> | ||
| + | |||
| + | |||
| + | </ul> | ||
| + | </li> | ||
| + | <li class="nav-item dropdown-box" style="background-color:darkslateblue;opacity: 0.9;"> | ||
| + | <a class="nav-link" href="#">Parts</a> | ||
| + | <ul class="dropdown-list"> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Contribution">Contribution</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Improve">Improve</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Newparts">New Parts</a></li> | ||
| + | |||
| + | </ul> | ||
| + | </li> | ||
| + | <li class="nav-item dropdown-box" style="background-color:indianred;opacity: 0.9;"> | ||
| + | <a class="nav-link" href="#">Human</a> | ||
| + | <ul class="dropdown-list"> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Human_Practices">Human Practices</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Education">Education</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Collaborations">Collaborations</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Partnership">Partnership</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Safety">Safety</a></li> | ||
| + | |||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Implementation">Implementation</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Entrepreneurship">Entrepreneurship</a></li> | ||
| + | |||
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</div> | </div> | ||
| − | </nav> | + | </div> |
| − | + | </nav> | |
| + | <!-- Navigation --> | ||
| − | + | <!-- Mobile Menu Start --> | |
| − | + | <nav class="mobile_menu hidden d-none"> | |
| − | + | <a href="https://2020.igem.org/Team:ZJU-China"><img class="mobile-logo" src="https://static.igem.org/mediawiki/2020/7/7f/T--ZJU-China--temt.png" alt="Logo"></a> | |
| − | + | <ul class="nav navbar-nav navbar-right menu"> | |
| − | + | <li class="dropdown active"> | |
| − | + | <a>Home</a> | |
| − | + | ||
| − | + | </li> | |
| − | + | <li class="dropdown"> | |
| − | + | <a>Team</a> | |
| − | + | <ul class="sub_menu"> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Team">Members</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Attributions">Attributions</a></li> | |
| − | </ | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Medals">Medals</a></li> |
| − | </li> | + | |
| − | + | </ul> | |
| − | + | </li> | |
| − | + | <li class="dropdown"> | |
| − | + | <a>Project</a> | |
| − | + | <ul class="sub_menu"> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Description">Description</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Background">Background</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Design">Design</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Magnetosome">Magnetosome</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Experiments">Experiments</a></li> | |
| − | </li> | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Results">Results</a></li> |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Future">Future</a></li> | |
| − | + | </ul> | |
| − | + | </li> | |
| − | + | <li class="dropdown"> | |
| − | + | <a>Model</a> | |
| − | + | <ul class="sub_menu"> | |
| − | </li> | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Model">Model</a></li> |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Engineering">Engineering</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Proof_Of_Concept">Proof of Concept</a></li> | |
| − | + | ||
| − | + | </ul> | |
| − | + | </li> | |
| − | + | <li class="dropdown"> | |
| − | + | <a>Parts</a> | |
| − | + | <ul class="sub_menu"> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Contribution">Contribution</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Improve">Improve</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Newparts">New Parts</a></li> | |
| − | + | ||
| − | + | </ul> | |
| − | </ | + | </li> |
| − | + | <li class="dropdown"> | |
| − | + | <a>Human</a> | |
| − | + | <ul class="sub_menu"> | |
| − | < | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Human_Practices">Human Practices</a></li> |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Education">Education</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Collaborations">Collaborations</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Partnership">Partnership</a></li> | |
| − | + | <li><a href="https://2020.igem.org/Team:ZJU-China/Safety">Safety</a></li> | |
| + | |||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Implementation">Implementation</a></li> | ||
| + | <li><a href="https://2020.igem.org/Team:ZJU-China/Entrepreneurship">Entrepreneurship</a></li> | ||
| + | |||
| + | </ul> | ||
| + | </li> | ||
| + | |||
| + | |||
| + | </ul> | ||
| + | </nav> | ||
| + | <!-- Mobile Menu End --> | ||
| + | </section> | ||
| + | <!--Main Menu/ Mobile Menu Section--> | ||
| − | + | <div id="swrapper"> | |
| − | + | <div id="mcontent" > | |
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<div class="pagename"> | <div class="pagename"> | ||
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| − | <div class="pagestyle" style="background-image: url('https://static.igem.org/mediawiki/2020/c/cd/T--ZJU-China--wiki_navback.jpg') | + | <div class="pagestyle" |
| + | style="background-image: url('https://static.igem.org/mediawiki/2020/c/cd/T--ZJU-China--wiki_navback.jpg');"> | ||
<div class="section Cloning" id="Cloning"> | <div class="section Cloning" id="Cloning"> | ||
<div class="container1"> | <div class="container1"> | ||
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<p> | <p> | ||
| − | When we had already chosen to develop a project that | + | When we had already chosen to develop a project that aimed to diagnose HER2 positive breast |
| − | + | cancer, we soon realized that MagHER2some was designed for patients in urgent | |
| − | MagHER2some as a accountable product to users and the whole society, we considered and evaluated each safety and ethical | + | conditions, and we should take strict safety measures to ensure our final product is qualified. |
| − | ZJU-China 2019 was developed a project | + | To aviod provoking tensions between doctor-patient relationships and make |
| − | + | MagHER2some as a accountable product to users and the whole society, we considered and evaluated | |
| − | + | each safety and ethical issues that might happen in real world. | |
| − | Browse following contents of this page for more safety & ethics details. You can also move to Human Practice Page to figure out how we conducted our Human | + | <br> |
| + | <br> | ||
| + | ZJU-China 2019 was developed a project focusing on female physical problems, either. The | ||
| + | comprehensive discussion on bioethics and safety that they conducted last year provided | ||
| + | useful guidance to us. As they mentioned before, it was of high importance for us to concentrate | ||
| + | on self-monitoring and ethical counseling. For this reason, we considered about | ||
| + | possible risks respecting laboratory, environment, clinical application and society. | ||
| + | <br> | ||
| + | <br> | ||
| + | Browse following contents of this page for more safety & ethics details. You can also move to | ||
| + | Human Practice Page to figure out how we conducted our Human Practice activities | ||
with our consideration of safety and ethics. | with our consideration of safety and ethics. | ||
</p> | </p> | ||
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<h2>Laboratory Safety</h2> | <h2>Laboratory Safety</h2> | ||
<p> | <p> | ||
| − | Working with living modified organisms is always risky. All standard precautions must be taken. Before starting to work in the lab, every member of our team was required to | + | Working with living modified organisms is always risky. All standard precautions must be taken. |
| − | attend the laboratory safety lesson and take a biosafety examination. Moreover, we have received extensive guidance and supervision regarding all our activities took place in | + | Before starting to work in the lab, every member of our team was required to |
| − | our laboratory, including study, discussion and equipment usage by our team PI, Prof. Ming Chen and experimental instructor, Associate Prof. Fan Yang to ensure the laboratory | + | attend the laboratory safety lesson and take a biosafety examination. Moreover, we have received |
| + | extensive guidance and supervision regarding all our activities took place in | ||
| + | our laboratory, including study, discussion and equipment usage by our team PI, Prof. Ming Chen | ||
| + | and experimental instructor, Associate Prof. Fan Yang to ensure the laboratory | ||
safety. | safety. | ||
</p> | </p> | ||
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<br> | <br> | ||
<p> | <p> | ||
| − | <b>Wearing safety：</b>Everyone wears protective lab coats, gloves and safety glasses before laboratory activities. Besides, due to the epidemic situation of COVID-19, everyone | + | <b>Wearing safety：</b>Everyone wears protective lab coats, gloves and safety glasses before |
| − | must wear a mask as | + | laboratory activities. Besides, due to the epidemic situation of COVID-19, everyone |
| + | must wear a mask as in the lab. | ||
</p> | </p> | ||
| + | <br> | ||
| + | <br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
| − | <img src="https://static.igem.org/mediawiki/2020/f/f7/T--ZJU-China--Safety_fig1.png" alt=""> | + | <img src="https://static.igem.org/mediawiki/2020/f/f7/T--ZJU-China--Safety_fig1.png" alt="" |
| − | <h6> | + | style="max-width: 55%;"> |
| + | <h6>Figure 1. Drawer specially for gloves and masks.</h6> | ||
</div> | </div> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
| − | <img src="https://static.igem.org/mediawiki/2020/d/db/T--ZJU-China--Safety_fig2.png" alt=""> | + | <img src="https://static.igem.org/mediawiki/2020/d/db/T--ZJU-China--Safety_fig2.png" alt="" |
| − | <h6> | + | style="max-width: 55%;"> |
| + | <h6>Figure 2. Door hanging our protective lab coats.</h6> | ||
</div> | </div> | ||
<p> | <p> | ||
| − | <b>Storage safety:</b>The chemical and biological reagents are stored in the proper place and suitable temperature. Hazardous or corrosive chemicals are stored separately. | + | <b>Storage safety: </b>The chemical and biological reagents are stored in the proper place and |
| + | suitable temperature. Hazardous or corrosive chemicals are stored separately. | ||
</p> | </p> | ||
| + | <br> | ||
| + | <br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
<img src="https://static.igem.org/mediawiki/2020/5/5d/T--ZJU-China--Safety_fig.3-5.png" alt=""> | <img src="https://static.igem.org/mediawiki/2020/5/5d/T--ZJU-China--Safety_fig.3-5.png" alt=""> | ||
| − | <h6> | + | <h6>Figure 3–5. Storage for chemical and biological reagents, students should record every time |
| + | when using chemicals.</h6> | ||
</div> | </div> | ||
| − | <p><b>Fire and electricity safety:</b> The temperature of the heater should not be set over 50℃ and the there must be at least one student in the lab when the heater is working. | + | <p><b>Fire and electricity safety:</b> The temperature of the heater should not be set over 50℃ and |
| + | the there must be at least one student in the lab when the heater is working. | ||
And the power should be turned off before the last one leaves the lab every day.</p> | And the power should be turned off before the last one leaves the lab every day.</p> | ||
| + | <br><br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
| − | <img src="https://static.igem.org/mediawiki/2020/7/70/T--ZJU-China--Safety_fig6.png" alt=""> | + | <img src="https://static.igem.org/mediawiki/2020/7/70/T--ZJU-China--Safety_fig6.png" alt="" |
| − | <h6> | + | style="max-width: 70%;"> |
| + | <h6>Figure 6. Power switch and heater.</h6> | ||
</div> | </div> | ||
| Line 353: | Line 477: | ||
<p> | <p> | ||
| − | <b>Safety Apparatus:</b> Our iGEM lab is equipped with safety apparatus such as fire extinguishers, emergency showers, eyewash stations, and first aid kits in case of dire | + | <b>Safety Apparatus:</b> Our iGEM lab is equipped with safety apparatus such as fire |
| + | extinguishers, emergency showers, eyewash stations, and first aid kits in case of dire | ||
situations. | situations. | ||
</p> | </p> | ||
| + | <br><br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
<img src="https://static.igem.org/mediawiki/2020/1/1e/T--ZJU-China--Safety_fig7-8.png" alt=""> | <img src="https://static.igem.org/mediawiki/2020/1/1e/T--ZJU-China--Safety_fig7-8.png" alt=""> | ||
| − | <h6> | + | <h6>Figure 7&8. The fire blanket and the fire extinguisher.</h6> |
</div> | </div> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
<img src="https://static.igem.org/mediawiki/2020/4/4e/T--ZJU-China--Safety_fig9-10.png" alt=""> | <img src="https://static.igem.org/mediawiki/2020/4/4e/T--ZJU-China--Safety_fig9-10.png" alt=""> | ||
| − | <h6> | + | <h6>Figure 9&10. The eyewash equipment and the emergency shower.</h6> |
</div> | </div> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
| − | <img src="https://static.igem.org/mediawiki/2020/a/ae/T--ZJU-China--Safety_fig11.png" alt=""> | + | <img src="https://static.igem.org/mediawiki/2020/a/ae/T--ZJU-China--Safety_fig11.png" alt="" |
| − | <h6> | + | style="max-width: 55%;"> |
| + | <h6>Figure 11. The first-aid case.</h6> | ||
</div> | </div> | ||
<p> | <p> | ||
<b>Waste disposal safety: </b> | <b>Waste disposal safety: </b> | ||
</p> | </p> | ||
| + | <br><br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
| − | <img src="https://static.igem.org/mediawiki/2020/2/27/T--ZJU-China--Safety_fig12.png" alt=""> | + | <img src="https://static.igem.org/mediawiki/2020/2/27/T--ZJU-China--Safety_fig12.png" alt="" |
| − | <h6> | + | style="max-width: 55%;"> |
| + | <h6>Figure 12. Flow chart of waste disposal.</h6> | ||
</div> | </div> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
<img src="https://static.igem.org/mediawiki/2020/1/1c/T--ZJU-China--Safety_fig13-14.png" alt=""> | <img src="https://static.igem.org/mediawiki/2020/1/1c/T--ZJU-China--Safety_fig13-14.png" alt=""> | ||
| − | <h6> | + | <h6>Figure 13&14. Garbage classification.</h6> |
</div> | </div> | ||
<p> | <p> | ||
| − | <b>New safety measures under the epidemic situation of COVID-19:</b> Due to the epidemic situation of COVID-19, all laboratories of Zhejiang University had strengthened the measures to prevent and control the virus. QR scanning and registration signing every day provided | + | <b>New safety measures under the epidemic situation of COVID-19:</b> Due to the epidemic |
| + | situation of COVID-19, all laboratories of Zhejiang University had strengthened the | ||
| + | measures to prevent and control the virus. QR scanning and registration signing every day | ||
| + | provided double entry records to track the members of ZJU-iGEM. And as we mentioned above, | ||
| + | masks must be worn when in the lab. | ||
</p> | </p> | ||
| + | <br> | ||
| + | <br> | ||
<div class="imgbox"> | <div class="imgbox"> | ||
<img src="https://static.igem.org/mediawiki/2020/d/d3/T--ZJU-China--Safety_fig15-17.png" alt=""> | <img src="https://static.igem.org/mediawiki/2020/d/d3/T--ZJU-China--Safety_fig15-17.png" alt=""> | ||
| − | <h6> | + | <h6>Figure 15-17. QR canning and signing for lab entry record.</h6> |
</div> | </div> | ||
| Line 395: | Line 530: | ||
<br> | <br> | ||
<p> | <p> | ||
| − | As iGEM committee emphasized, we have paid great attention to the possible risks of MagHER2some during our entire process of iGEM. And we have complied with relevant | + | As iGEM committee emphasized, we have paid great attention to the possible risks of MagHER2some |
| − | regulations and completed the filling of safety forms and animal use form carfully a month earlier. | + | during our entire process of iGEM. And we have complied with relevant |
| + | regulations and completed the filling of safety forms and animal use form carfully a month | ||
| + | earlier. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| Line 404: | Line 542: | ||
<h3>Cell-free system</h3> | <h3>Cell-free system</h3> | ||
<p> | <p> | ||
| − | The production and release of genetically modified organisms (GMOs) are controversial topics within present scientific community<a href="# | + | The production and release of genetically modified organisms (GMOs) are controversial topics |
| − | also considered about the possibility risks may cause by two chassis organisms <i | + | within present scientific community<a href="#references"><sup>[1]</sup></a>, thus we |
| − | organisms used in MagHER2some. In accordance with | + | also considered about the possibility risks may cause by two chassis organisms, |
| + | <i>Magnetospirillum gryphiswaldense</i> and <i>E.coli</i> SHuffle<sup>®</sup> and other | ||
| + | organisms used in MagHER2some. In accordance with iGEM safety rules and standards, we do not | ||
| + | plan to release any GMOs actively in any cases to strictly aviod possible | ||
ecological disorders happening. | ecological disorders happening. | ||
| − | Moreover, in accordance with | + | Moreover, in accordance with iGEM safety rules and standards, our final product is based on |
| − | design not only meets | + | native magnetosome and created a cell-free system instead of alive bacteria. This |
| + | design not only meets "no-release" principle perfectly, but also minimizes possible cytotoxicity | ||
| + | and immunotoxicity caused by bacteria when our novel MRI contrast agent is | ||
injected into our users. | injected into our users. | ||
| − | What needs to be added here is the biotools we implemented on MagHER2some are all protein and without any nucleic acid, which means MagHER2some is totally nonhazardous and | + | What needs to be added here is that the biotools we implemented on MagHER2some are all protein |
| − | unable to express any gene in all circumstances, especially when it is released into environment by accdient or injected into patients' circulatory system. | + | and without any nucleic acid, which means MagHER2some is totally nonhazardous and |
| + | unable to express any gene in all circumstances, especially when it is released into environment | ||
| + | by accdient or injected into patients' circulatory system. | ||
</p> | </p> | ||
<br> | <br> | ||
| Line 421: | Line 566: | ||
</p> | </p> | ||
<p> | <p> | ||
| − | The main reason why we chose | + | The main reason why we chose magnetosomes as the most basic BioBricks is that they have been |
| − | reported that chemical toxicity of magnetosome from iron ions is | + | proven to be highly biocompatible<a href="#references"><sup>[2]</sup></a>. It has been |
| − | primary safety issues of magnetosome may be relevant to (a) their nanoscale size, which leads to embolism, blockage, and deposition in the body and (b) impurities, particularly | + | reported that chemical toxicity of magnetosome from iron ions is negligible<a |
| − | proteins, nucleic acids, and polysaccharides associated with magnetosome extracted from bacterial cells, resulting in immunotoxicity. | + | href="#references"><sup>[3][4]</sup></a> due to the insolubility of |
| + | Fe<sub>3</sub>O<sub>4</sub>. Therefore, the | ||
| + | primary safety issues of magnetosome may be relevant to (a) their nanoscale size, which leads to | ||
| + | embolism, blockage, and deposition in the body and (b) impurities, particularly | ||
| + | proteins, nucleic acids, and polysaccharides associated with magnetosome extracted from | ||
| + | bacterial cells, resulting in immunotoxicity. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | As we had a further understanding of those studies related to the biocompatibility of | + | As we had a further understanding of those studies related to the biocompatibility of |
| − | and host tissue elimination following administration of | + | magnetosomes, we found out that the existing studies have shown the body tissue distribution |
| − | fibroblasts<a href="# | + | and host tissue elimination following administration of magnetosomes into the vascular system<a |
| + | href="#references"><sup>[5]</sup></a> and <i>in vitro</i> cytotoxicity for mouse | ||
| + | fibroblasts<a href="#references"><sup>[6]</sup></a>. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | It has been observed that | + | It has been observed that magnetosomes displayed targeted distribution in Sprague DawleySD rat |
| − | congestion or infarction in vivo | + | liver except other organs, suggesting that magnetosomes may avoid incurring organ |
| + | congestion or infarction <i>in vivo</i> relyed on the lower likelihood of congregation than | ||
| + | other nanoparticles. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | Purified and sterilized magnetosomes were found to be nontoxic to mouse fibroblasts in vitro. Moreover, another research shows that the injection of 1 mg | + | Purified and sterilized magnetosomes were found to be nontoxic to mouse fibroblasts <i>in |
| − | increase body temperature of rabbits during the pyrogen test, which showed antigens or pyrogens free with magnetosome administration <a href="# | + | vitro</i>. Moreover, another research shows that the injection of 1 mg magnetosomes did not |
| + | increase the body temperature of rabbits during the pyrogen test, which showed antigens or | ||
| + | pyrogens free with magnetosome administration<a href="#references"><sup>[7]</sup></a>. | ||
</p> | </p> | ||
| Line 447: | Line 606: | ||
</p> | </p> | ||
<p> | <p> | ||
| − | ZZ domain is a homologous immunoglobulin (Ig)-binding domain which belongs to <i>Staphy- lococcus aureus</i> | + | ZZ domain is a homologous immunoglobulin (Ig)-binding domain which belongs to <i>Staphy- |
| − | created a recombinant protein mamC-ZZ which | + | lococcus aureus</i> protein A (SpA). According to our biological engineering, we |
| − | antibody-binding BioBricks, and SpA which ZZ domain belongs | + | created a recombinant protein mamC-ZZ which was able to bind to the fragment crystallizable (Fc) |
| + | region of single chain antibody fragment (scFv) tightly. ZZ domain is a reactive | ||
| + | antibody-binding BioBricks, and SpA to which ZZ domain belongs is widely used in clinical | ||
| + | application<a href="#references"><sup>[8][9]</sup></a>, referring to a conclusion that ZZ | ||
domain is safe enough to be used as a BioBricks. | domain is safe enough to be used as a BioBricks. | ||
<p></p> | <p></p> | ||
<br> | <br> | ||
<p> | <p> | ||
| − | <b>C. | + | <b>C. Single chain antibody fragment (scFv)</b> |
</p> | </p> | ||
<p> | <p> | ||
| − | Single chain antibody fragment (scFv) is the most significant important part of our biological engineering design that determines diagnostic efficiency and imaging results of | + | Single chain antibody fragment (scFv) is the most significant important part of our biological |
| − | + | engineering design that determines diagnostic efficiency and imaging results of | |
| − | with high affinity to the extracellular domain of HER2 in a high affinity way. Trastuzumab is a relatively safe and usually well-tolerated drug<a | + | MagHER2some. Accordingly, we chose a certain kind of scFv modified from a widely used breast |
| − | + | cancer medicine Trastuzumab, which had the ability to bind | |
| − | href="# | + | with high affinity to the extracellular domain of HER2 in a high affinity way. Trastuzumab is a |
| − | relating to the homeostatic regulation of endogenous | + | relatively safe and usually well-tolerated drug<a href="#references"><sup>[10]</sup></a>. As a |
| − | with unwanted targets (frequently one of the main causes of | + | monoclonaln antibody, trastuzumab is not modulated by the classic routes of drug elimination<a |
| + | href="#references"><sup>[11]</sup></a>. Its clearance does not appear to occur or excretion | ||
| + | and liver metabolism as conventional drugs, but in an analogue mechanism | ||
| + | relating to the homeostatic regulation of endogenous immunoglobulin G<a | ||
| + | href="#references"><sup>[12]</sup></a>. Thus, problem such as drug–drug interactions and | ||
| + | interference | ||
| + | with unwanted targets (frequently one of the main causes of drugs' 'adverse effects') is | ||
| + | unlikely. | ||
</p> | </p> | ||
<br> | <br> | ||
<p> | <p> | ||
| − | <b>Final Product: reflects biosafety as a whole </b> | + | <b>D. Final Product: reflects biosafety as a whole </b> |
<br> | <br> | ||
| − | Those studies that mentioned above are all strong evidence which ensure all BioBricks of MagHER2some to be safe enough. However, there is still a long term exploration for us | + | Those studies that mentioned above are all strong evidence which ensure all BioBricks of |
| + | MagHER2some to be safe enough. However, there is still a long term exploration for us | ||
to implement our biological engineering into a reliable final product. | to implement our biological engineering into a reliable final product. | ||
</p> | </p> | ||
| Line 477: | Line 647: | ||
<p> | <p> | ||
| − | <b>Interacrtion between BioBricks:</b>We carried out a series of protein three-dimensional | + | <b>Interacrtion between BioBricks: </b>We carried out a series of predicted protein |
| − | mamC-ZZ and Trastuzumab. No evidence | + | three-dimensional structures about the results of the interaction between recombinant protein |
| + | mamC-ZZ and Trastuzumab. No evidence showed that there had any effective domain been changed or | ||
| + | blocked because of interaction, referred MagHER2some to a therapeutic beneficial | ||
product. | product. | ||
</p> | </p> | ||
| Line 485: | Line 657: | ||
<p> | <p> | ||
<b>Suitable Nanoscale: </b> | <b>Suitable Nanoscale: </b> | ||
| − | We decided to control the size of MagHER2some | + | We decided to control the size of MagHER2some in a appopriate range (15–35 nm), which not only |
| − | also | + | prevented that MagHER2some would be removed by reticuloendothelial system, but |
| + | also ensured that MagHER2some can be excreted into urine and feces <a | ||
| + | href="#references"><sup[13][14]</sup></a>. | ||
</p> | </p> | ||
<br> | <br> | ||
| Line 494: | Line 668: | ||
Further Clinical Trials: | Further Clinical Trials: | ||
</b> | </b> | ||
| − | What | + | What need to be mentioned again is that we still need a long term exploration on biosafety. We |
| − | drug instruction.You can see more clinical trials design here | + | will conduct further clinical trials of MagHER2some and launch the corresponding |
| + | drug instruction. You can see more clinical trials design here at <a | ||
| + | href="https://2020.igem.org/Team:ZJU-China/Implementation#technical_support" | ||
| + | class="outerlink" style="color:#0a8aff;">LINK</a>. | ||
| + | <br> | ||
</p> | </p> | ||
<p> | <p> | ||
| − | Finally, we | + | Finally, we plan to optimize the producing procedure of the final products, which will be |
| + | designed into nonhazardous materials and harmless reagents. | ||
</p> | </p> | ||
| Line 511: | Line 690: | ||
<h2>Bioethics</h2> | <h2>Bioethics</h2> | ||
<p> | <p> | ||
| − | We have controlled possible risks from two | + | We have controlled possible risks from two perspectives: the laboratory safety and the |
| − | + | biological | |
| + | engineering safety. However, there are still many potential ethical risks need to be | ||
| + | prejudged. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | + | "Discussions of bioethical challenges take place in the media, in the academy, in classrooms, | |
| − | just for us. From another perspective, bioethics can be considered in terms of | + | but also in labs, offices, and hospital wards." Problem in bioethics is not |
| + | just for us. From another perspective, bioethics can be considered in terms of human, animals | ||
| + | (experimental organisms) and the environment. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | All ethical safety considerations of | + | All ethical safety considerations of MagHER2some are discussed as below. |
</p> | </p> | ||
<br> | <br> | ||
| Line 529: | Line 714: | ||
<br> | <br> | ||
<p> | <p> | ||
| − | <b>Experimental Animal | + | <b>Use of Experimental Animal:</b> |
| − | As the very beginning step of bioethics consideration, we have carried on the comprehensive discussion about this part through our animal use form | + | As the very beginning step of bioethics consideration, we have carried on the comprehensive |
| + | discussion about this part through our animal use form. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | Moreover, we | + | Moreover, we were reminded that some contents of our animal use form still needed to be |
| − | not yet been obtained at that time. Now, in order to | + | improved, because most of the <i>in vitro</i> experiments of MagHER2some had |
| − | from in vitro experiments, and | + | not yet been obtained at that time. Now, in order to supply and clarify the contents of |
| + | animal work, we have redesigned the animal experiments based on the results obtained | ||
| + | from <i>in vitro</i> experiments, and focused on animal welfare more detailly. Please see the | ||
| + | file below for details. | ||
| + | <br> | ||
| + | <a href="https://static.igem.org/mediawiki/2020/b/bf/T--ZJU-China--Improved_animal_experiment_plan.pdf" | ||
| + | class="outerlink" style="color:#0a8aff;">Improved Animal Experiment Plan.pdf</a> | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | <b>Commercial Matters:</b> We are devoted to bring MagHER2some into medical market, and hope it | + | <b>Commercial Matters:</b> We are devoted to bring MagHER2some into medical market, and hope it |
| − | at | + | can benefit both doctors and patients. More commercial risks |
| + | at <a href="https://2020.igem.org/Team:ZJU-China/Implementation" class="outerlink" style="color:#0a8aff;">HERE</a>. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | <b>Public Engagement:</b> | + | <b>Public Engagement:</b> We believe that our sponsors, taxpayers have |
| − | projects | + | the right to know and to participate in, you can see how the public participated in our |
| + | projects through our promotion activities in cooperation with museum and the Red Cross at <a href="https://2020.igem.org/Team:ZJU-China/Education" class="outerlink" style="color:#0a8aff;">LINK</a>, | ||
| + | and how our project is responsible for the public such as protecting | ||
| + | the personal portrait right of patients. | ||
</p> | </p> | ||
<br> | <br> | ||
| Line 556: | Line 755: | ||
<p> | <p> | ||
<b>Patent and Intellectual Property Protection:</b> | <b>Patent and Intellectual Property Protection:</b> | ||
| − | + | We plan to patent MagHER2some after iGEM. Therefore, we have already consulted a | |
| + | law firm for relevant issues. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | <b> | + | <b>Strict Supplement:</b> |
| − | Considering | + | Considering biosequrity, we only provide the final products to certified medical institutions. |
| − | our products, | + | In this way, the possibility of human error which people may misuse or abuse |
| + | our products, will be minimized. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
<b>Product Instructions:</b> | <b>Product Instructions:</b> | ||
| − | In terms of a thorough biosafety guarantee, we are going to undertake further clinical trials and upload a informative instructions to illustrate the right execution to | + | In terms of a thorough biosafety guarantee, we are going to undertake further clinical trials |
| − | doctor who will use MagHER2some in future. More details of clinical trials at | + | and upload a informative instructions to illustrate the right execution to each |
| + | doctor who will use MagHER2some in future. More details of clinical trials at <a | ||
| + | href="https://2020.igem.org/Team:ZJU-China/Implementation" class="outerlink" style="color:#0a8aff;">HERE</a>. | ||
</p> | </p> | ||
| Line 582: | Line 787: | ||
<h2>Always be Responsible</h2> | <h2>Always be Responsible</h2> | ||
<p> | <p> | ||
| − | We are ready to use | + | We are ready to use MagHER2some as a product to prepare for future |
| − | real world. In terms of ethics, China is different from foreign countries. | + | commercialization and to face various challenges that may come out in the |
| − | laboratory. This is also a reason that we | + | real world. In terms of ethics, China is different from foreign countries. Similar to many |
| + | Chinese iGEM | ||
| + | teams, we do not have systematic bioethics learning before entering the | ||
| + | laboratory. This is also a reason that we did not achieve a thoroughly ethical consideration of | ||
| + | experimental animals before. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | However, once MagHER2some is in clinical trials involving human | + | However, once MagHER2some is in clinical trials involving human, a new set of |
| − | participants to guarantee their participation is voluntary and informed. That means we need take | + | challenges which are from ensuring informed consent to protecting vulnerable research |
| − | only | + | participants to guarantee their participation will arise, which is voluntary and informed. That |
| + | means we need to take more strict ethical measures. Even though the further clinical trials are | ||
| + | only in design stage, we still need to ensure whether the plan is ethical enough. | ||
</p> | </p> | ||
| + | <br> | ||
<p> | <p> | ||
| − | Once again, there is still a long term exploration for us to see the benefits of MagHER2some. As an iGEM team, we need to pay more attention to self-monitoring, raise awareness | + | Once again, there is still a long term exploration for us to see the benefits of MagHER2some. As |
| − | of ethical counseling | + | an iGEM team, we need to pay more attention to self-monitoring, raise awareness |
| + | of ethical counseling and ask boldly before we achieve our goal. | ||
</p> | </p> | ||
<br> | <br> | ||
| Line 603: | Line 817: | ||
| − | <div class="section our-team" id=" | + | <div class="section our-team" id="references"> |
<div class="container1"> | <div class="container1"> | ||
| − | <h2> | + | <h2>References</h2> |
<p> | <p> | ||
| − | [1] Ledford H. CRISPR conundrum: strict European court ruling leaves food-testing labs without a plan[J]. Nature, 2019, 572(7767): 15-16. | + | [1]. Ledford H. CRISPR conundrum: strict European court ruling leaves food-testing labs without a |
| + | plan[J]. <i>Nature, 2019</i>, 572(7767): 15-16. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [2] Sun J, Li Y, Liang XJ, Wang PC. Bacterial Magnetosome: A Novel Biogenetic Magnetic Targeted Drug Carrier with Potential Multifunctions. J Nanomater. | + | [2]. Sun J, Li Y, Liang XJ, Wang PC. Bacterial Magnetosome: A Novel Biogenetic Magnetic Targeted |
| − | 2011;2011(2011):469031-469043. | + | Drug Carrier with Potential Multifunctions. <i>J Nanomater. |
| + | 2011</i>;2011(2011):469031-469043. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [3] Häfeli UO, Pauer GJ. In vitro and in vivo toxicity of magnetic microspheres. | + | [3]. Häfeli UO, Pauer GJ. <i>In vitro</i> and <i>in vivo</i> toxicity of magnetic microspheres. |
| + | <i>Journal of Magnetism and Magnetic Materials. 1999</i>;vol. 194(no. 1):76–82. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [4] | + | [4]. Wagner V, Dullaart A, Bock AK, Zweck A. The emerging nanomedicine landscape. <i>Nature |
| + | Biotechnology.</i> 2006;vol. 24(no. 10):1211–1217. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [5] Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy | + | [5]. Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial |
| − | Sprague-Dawley rats. J Nanosci Nanotechnol. 2009 Mar;9(3):1881-5. | + | magnetosomes isolated from <i>Magnetospirillum gryphiswaldense</i> MSR-1 in healthy |
| + | Sprague-Dawley rats. <i>J Nanosci Nanotechnol. 2009</i> Mar;9(3):1881-5. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [6] Xiang L, Wei J, Jianbo S, Guili W, Feng G, Ying L. Purified and sterilized magnetosomes from Magnetospirillum gryphiswaldense MSR-1 were not toxic to mouse fibroblasts in | + | [6]. Xiang L, Wei J, Jianbo S, Guili W, Feng G, Ying L. Purified and sterilized magnetosomes from |
| − | vitro. Lett Appl Microbiol. 2007 Jul;45(1):75-81. | + | <i>Magnetospirillum gryphiswaldense</i> MSR-1 were not toxic to mouse fibroblasts in |
| + | vitro. <i>Lett Appl Microbiol. 2007</i> Jul;45(1):75-81. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [7] Sun J, Tang T, Duan J, Xu PX, Wang Z, Zhang Y, Wu L, Li Y. Biocompatibility of bacterial magnetosomes: acute toxicity, immunotoxicity and cytotoxicity. Nanotoxicology. 2010 | + | [7]. Sun J, Tang T, Duan J, Xu PX, Wang Z, Zhang Y, Wu L, Li Y. Biocompatibility of bacterial |
| + | magnetosomes: acute toxicity, immunotoxicity and cytotoxicity. <i>Nanotoxicology. 2010</i> | ||
Sep;4(3):271-83. | Sep;4(3):271-83. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [8] Navegantes, K. C., | + | [8]. Navegantes, K. C., Gomes, R. S., Pereira, P. A. T., Czaikoski, P. G., Azevedo, C. H. M., and |
| + | Monteiro, M. C. (2017) J. <i>Transl. Med. 15</i>, 36. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [9] Idowu, O., | + | [9]. Idowu, O., and Heading, K. (2018) J. Small Anim. <i>Pract. 59</i>, 183–187. |
</p> | </p> | ||
<p> | <p> | ||
| − | [10] Senkus E, Kyriakides S, Penault-Llorca F, Poortmans P, Thompson A, Zackrisson S, et al. Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment | + | [10]. Senkus E, Kyriakides S, Penault-Llorca F, Poortmans P, Thompson A, Zackrisson S, et al. |
| − | and follow-up. Ann Oncol. 2013;24(Suppl 6):7–23. | + | Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment |
| + | and follow-up. <i>Ann Oncol. 2013</i>;24(Suppl 6):7–23. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [11] European Medicines Agency. Summary of product characteristics: | + | [11]. European Medicines Agency. Summary of product characteristics: |
| − | Herceptin.http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Product_Information/human/000278/WC500074922.pdf. Accessed 9 Feb 2016. | + | Herceptin.http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Product_Information/human/000278/WC500074922.pdf. |
| + | Accessed 9 Feb 2016. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [12] Azanza J-R, Sádaba B, Gómez-Guiu A. Monoclonal antibodies: pharmacokinetics as a basis for new dosage regimens? J Oncol Pharm Pract. 2015;21:370–6. | + | [12]. Azanza J-R, Sádaba B, Gómez-Guiu A. Monoclonal antibodies: pharmacokinetics as a basis for |
| + | new dosage regimens? <i>J Oncol Pharm Pract. 2015<i/>;21:370–6. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [13] Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy | + | [13]. Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial |
| − | Sprague-Dawley rats. J Nanosci Nanotechnol. 2009 Mar;9(3):1881-5. | + | magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy |
| + | Sprague-Dawley rats. <i>J Nanosci Nanotechnol. 2009</i> Mar;9(3):1881-5. | ||
</p> | </p> | ||
<p> | <p> | ||
| − | [14] | + | [14]. Meseret Amde, Jing-fu Liu, Zhi-Qiang Tan, Deribachew Bekana, Transformation and |
| − | review. Environmental Pollution. 2017;vol. 230:250-267. | + | bioavailability of metal oxide nanoparticles in aquatic and terrestrial environments. A |
| + | review. <i>Environmental Pollution. 2017</i>;vol. 230:250-267. | ||
</p> | </p> | ||
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Latest revision as of 03:55, 28 October 2020
Safety
Overview
When we had already chosen to develop a project that aimed to diagnose HER2 positive breast
cancer, we soon realized that MagHER2some was designed for patients in urgent
conditions, and we should take strict safety measures to ensure our final product is qualified.
To aviod provoking tensions between doctor-patient relationships and make
MagHER2some as a accountable product to users and the whole society, we considered and evaluated
each safety and ethical issues that might happen in real world.
ZJU-China 2019 was developed a project focusing on female physical problems, either. The
comprehensive discussion on bioethics and safety that they conducted last year provided
useful guidance to us. As they mentioned before, it was of high importance for us to concentrate
on self-monitoring and ethical counseling. For this reason, we considered about
possible risks respecting laboratory, environment, clinical application and society.
Browse following contents of this page for more safety & ethics details. You can also move to
Human Practice Page to figure out how we conducted our Human Practice activities
with our consideration of safety and ethics.
Laboratory Safety
Working with living modified organisms is always risky. All standard precautions must be taken. Before starting to work in the lab, every member of our team was required to attend the laboratory safety lesson and take a biosafety examination. Moreover, we have received extensive guidance and supervision regarding all our activities took place in our laboratory, including study, discussion and equipment usage by our team PI, Prof. Ming Chen and experimental instructor, Associate Prof. Fan Yang to ensure the laboratory safety.
Wearing safety：Everyone wears protective lab coats, gloves and safety glasses before laboratory activities. Besides, due to the epidemic situation of COVID-19, everyone must wear a mask as in the lab.
Figure 1. Drawer specially for gloves and masks.
Figure 2. Door hanging our protective lab coats.
Storage safety: The chemical and biological reagents are stored in the proper place and suitable temperature. Hazardous or corrosive chemicals are stored separately.
Figure 3–5. Storage for chemical and biological reagents, students should record every time when using chemicals.
Fire and electricity safety: The temperature of the heater should not be set over 50℃ and the there must be at least one student in the lab when the heater is working. And the power should be turned off before the last one leaves the lab every day.
Figure 6. Power switch and heater.
Safety Apparatus: Our iGEM lab is equipped with safety apparatus such as fire extinguishers, emergency showers, eyewash stations, and first aid kits in case of dire situations.
Figure 7&8. The fire blanket and the fire extinguisher.
Figure 9&10. The eyewash equipment and the emergency shower.
Figure 11. The first-aid case.
Waste disposal safety:
Figure 12. Flow chart of waste disposal.
Figure 13&14. Garbage classification.
New safety measures under the epidemic situation of COVID-19: Due to the epidemic situation of COVID-19, all laboratories of Zhejiang University had strengthened the measures to prevent and control the virus. QR scanning and registration signing every day provided double entry records to track the members of ZJU-iGEM. And as we mentioned above, masks must be worn when in the lab.
Figure 15-17. QR canning and signing for lab entry record.
Insurance of Biosafety
As iGEM committee emphasized, we have paid great attention to the possible risks of MagHER2some during our entire process of iGEM. And we have complied with relevant regulations and completed the filling of safety forms and animal use form carfully a month earlier.
In this page, the main safety issues of our project design are discussed below.
Cell-free system
The production and release of genetically modified organisms (GMOs) are controversial topics within present scientific community[1], thus we also considered about the possibility risks may cause by two chassis organisms, Magnetospirillum gryphiswaldense and E.coli SHuffle® and other organisms used in MagHER2some. In accordance with iGEM safety rules and standards, we do not plan to release any GMOs actively in any cases to strictly aviod possible ecological disorders happening. Moreover, in accordance with iGEM safety rules and standards, our final product is based on native magnetosome and created a cell-free system instead of alive bacteria. This design not only meets "no-release" principle perfectly, but also minimizes possible cytotoxicity and immunotoxicity caused by bacteria when our novel MRI contrast agent is injected into our users. What needs to be added here is that the biotools we implemented on MagHER2some are all protein and without any nucleic acid, which means MagHER2some is totally nonhazardous and unable to express any gene in all circumstances, especially when it is released into environment by accdient or injected into patients' circulatory system.
BioBricks
A. Magnetosome
The main reason why we chose magnetosomes as the most basic BioBricks is that they have been proven to be highly biocompatible[2]. It has been reported that chemical toxicity of magnetosome from iron ions is negligible[3][4] due to the insolubility of Fe3O4. Therefore, the primary safety issues of magnetosome may be relevant to (a) their nanoscale size, which leads to embolism, blockage, and deposition in the body and (b) impurities, particularly proteins, nucleic acids, and polysaccharides associated with magnetosome extracted from bacterial cells, resulting in immunotoxicity.
As we had a further understanding of those studies related to the biocompatibility of magnetosomes, we found out that the existing studies have shown the body tissue distribution and host tissue elimination following administration of magnetosomes into the vascular system[5] and in vitro cytotoxicity for mouse fibroblasts[6].
It has been observed that magnetosomes displayed targeted distribution in Sprague DawleySD rat liver except other organs, suggesting that magnetosomes may avoid incurring organ congestion or infarction in vivo relyed on the lower likelihood of congregation than other nanoparticles.
Purified and sterilized magnetosomes were found to be nontoxic to mouse fibroblasts in vitro. Moreover, another research shows that the injection of 1 mg magnetosomes did not increase the body temperature of rabbits during the pyrogen test, which showed antigens or pyrogens free with magnetosome administration[7].
B. ZZ domain
ZZ domain is a homologous immunoglobulin (Ig)-binding domain which belongs to Staphy- lococcus aureus protein A (SpA). According to our biological engineering, we created a recombinant protein mamC-ZZ which was able to bind to the fragment crystallizable (Fc) region of single chain antibody fragment (scFv) tightly. ZZ domain is a reactive antibody-binding BioBricks, and SpA to which ZZ domain belongs is widely used in clinical application[8][9], referring to a conclusion that ZZ domain is safe enough to be used as a BioBricks.
C. Single chain antibody fragment (scFv)
Single chain antibody fragment (scFv) is the most significant important part of our biological engineering design that determines diagnostic efficiency and imaging results of MagHER2some. Accordingly, we chose a certain kind of scFv modified from a widely used breast cancer medicine Trastuzumab, which had the ability to bind with high affinity to the extracellular domain of HER2 in a high affinity way. Trastuzumab is a relatively safe and usually well-tolerated drug[10]. As a monoclonaln antibody, trastuzumab is not modulated by the classic routes of drug elimination[11]. Its clearance does not appear to occur or excretion and liver metabolism as conventional drugs, but in an analogue mechanism relating to the homeostatic regulation of endogenous immunoglobulin G[12]. Thus, problem such as drug–drug interactions and interference with unwanted targets (frequently one of the main causes of drugs' 'adverse effects') is unlikely.
D. Final Product: reflects biosafety as a whole
Those studies that mentioned above are all strong evidence which ensure all BioBricks of
MagHER2some to be safe enough. However, there is still a long term exploration for us
to implement our biological engineering into a reliable final product.
Interacrtion between BioBricks: We carried out a series of predicted protein three-dimensional structures about the results of the interaction between recombinant protein mamC-ZZ and Trastuzumab. No evidence showed that there had any effective domain been changed or blocked because of interaction, referred MagHER2some to a therapeutic beneficial product.
Suitable Nanoscale:
We decided to control the size of MagHER2some in a appopriate range (15–35 nm), which not only
prevented that MagHER2some would be removed by reticuloendothelial system, but
also ensured that MagHER2some can be excreted into urine and feces
Further Clinical Trials:
What need to be mentioned again is that we still need a long term exploration on biosafety. We
will conduct further clinical trials of MagHER2some and launch the corresponding
drug instruction. You can see more clinical trials design here at LINK.
Finally, we plan to optimize the producing procedure of the final products, which will be designed into nonhazardous materials and harmless reagents.
Bioethics
We have controlled possible risks from two perspectives: the laboratory safety and the biological engineering safety. However, there are still many potential ethical risks need to be prejudged.
"Discussions of bioethical challenges take place in the media, in the academy, in classrooms, but also in labs, offices, and hospital wards." Problem in bioethics is not just for us. From another perspective, bioethics can be considered in terms of human, animals (experimental organisms) and the environment.
All ethical safety considerations of MagHER2some are discussed as below.
During iGEM
Use of Experimental Animal: As the very beginning step of bioethics consideration, we have carried on the comprehensive discussion about this part through our animal use form.
Moreover, we were reminded that some contents of our animal use form still needed to be
improved, because most of the in vitro experiments of MagHER2some had
not yet been obtained at that time. Now, in order to supply and clarify the contents of
animal work, we have redesigned the animal experiments based on the results obtained
from in vitro experiments, and focused on animal welfare more detailly. Please see the
file below for details.
Improved Animal Experiment Plan.pdf
Commercial Matters: We are devoted to bring MagHER2some into medical market, and hope it can benefit both doctors and patients. More commercial risks at HERE.
Public Engagement: We believe that our sponsors, taxpayers have the right to know and to participate in, you can see how the public participated in our projects through our promotion activities in cooperation with museum and the Red Cross at LINK, and how our project is responsible for the public such as protecting the personal portrait right of patients.
After iGEM
Patent and Intellectual Property Protection: We plan to patent MagHER2some after iGEM. Therefore, we have already consulted a law firm for relevant issues.
Strict Supplement: Considering biosequrity, we only provide the final products to certified medical institutions. In this way, the possibility of human error which people may misuse or abuse our products, will be minimized.
Product Instructions: In terms of a thorough biosafety guarantee, we are going to undertake further clinical trials and upload a informative instructions to illustrate the right execution to each doctor who will use MagHER2some in future. More details of clinical trials at HERE.
Always be Responsible
We are ready to use MagHER2some as a product to prepare for future commercialization and to face various challenges that may come out in the real world. In terms of ethics, China is different from foreign countries. Similar to many Chinese iGEM teams, we do not have systematic bioethics learning before entering the laboratory. This is also a reason that we did not achieve a thoroughly ethical consideration of experimental animals before.
However, once MagHER2some is in clinical trials involving human, a new set of challenges which are from ensuring informed consent to protecting vulnerable research participants to guarantee their participation will arise, which is voluntary and informed. That means we need to take more strict ethical measures. Even though the further clinical trials are only in design stage, we still need to ensure whether the plan is ethical enough.
Once again, there is still a long term exploration for us to see the benefits of MagHER2some. As an iGEM team, we need to pay more attention to self-monitoring, raise awareness of ethical counseling and ask boldly before we achieve our goal.
References
[1]. Ledford H. CRISPR conundrum: strict European court ruling leaves food-testing labs without a plan[J]. Nature, 2019, 572(7767): 15-16.
[2]. Sun J, Li Y, Liang XJ, Wang PC. Bacterial Magnetosome: A Novel Biogenetic Magnetic Targeted Drug Carrier with Potential Multifunctions. J Nanomater. 2011;2011(2011):469031-469043.
[3]. Häfeli UO, Pauer GJ. In vitro and in vivo toxicity of magnetic microspheres. Journal of Magnetism and Magnetic Materials. 1999;vol. 194(no. 1):76–82.
[4]. Wagner V, Dullaart A, Bock AK, Zweck A. The emerging nanomedicine landscape. Nature Biotechnology. 2006;vol. 24(no. 10):1211–1217.
[5]. Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy Sprague-Dawley rats. J Nanosci Nanotechnol. 2009 Mar;9(3):1881-5.
[6]. Xiang L, Wei J, Jianbo S, Guili W, Feng G, Ying L. Purified and sterilized magnetosomes from Magnetospirillum gryphiswaldense MSR-1 were not toxic to mouse fibroblasts in vitro. Lett Appl Microbiol. 2007 Jul;45(1):75-81.
[7]. Sun J, Tang T, Duan J, Xu PX, Wang Z, Zhang Y, Wu L, Li Y. Biocompatibility of bacterial magnetosomes: acute toxicity, immunotoxicity and cytotoxicity. Nanotoxicology. 2010 Sep;4(3):271-83.
[8]. Navegantes, K. C., Gomes, R. S., Pereira, P. A. T., Czaikoski, P. G., Azevedo, C. H. M., and Monteiro, M. C. (2017) J. Transl. Med. 15, 36.
[9]. Idowu, O., and Heading, K. (2018) J. Small Anim. Pract. 59, 183–187.
[10]. Senkus E, Kyriakides S, Penault-Llorca F, Poortmans P, Thompson A, Zackrisson S, et al. Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2013;24(Suppl 6):7–23.
[11]. European Medicines Agency. Summary of product characteristics: Herceptin.http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Product_Information/human/000278/WC500074922.pdf. Accessed 9 Feb 2016.
[12]. Azanza J-R, Sádaba B, Gómez-Guiu A. Monoclonal antibodies: pharmacokinetics as a basis for new dosage regimens? J Oncol Pharm Pract. 2015;21:370–6.
[13]. Sun JB, Wang ZL, Duan JH, Ren J, Yang XD, Dai SL, Li Y. Targeted distribution of bacterial magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy Sprague-Dawley rats. J Nanosci Nanotechnol. 2009 Mar;9(3):1881-5.
[14]. Meseret Amde, Jing-fu Liu, Zhi-Qiang Tan, Deribachew Bekana, Transformation and bioavailability of metal oxide nanoparticles in aquatic and terrestrial environments. A review. Environmental Pollution. 2017;vol. 230:250-267.
