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Laboratory Safety

The iGEM-Technion team's labs are in full compliance with the safety and security rules of both the iGEM competition and the Technion institute. Said laboratories are in the biotechnology and food engineering faculty (located inside the Technion institute) and are certified as Biosafety levels 1 and 2.

All team members received training and passed different safety tests provided by the Technion safety department prior to entering the lab and beginning our lab work. Additionally, all sorts of work which includes complicated methods, requiring special equipment or a special working area (such as tissue culture room) were done with an authorized supervisor.

Personal Safety

We are working with all the necessary safety equipment such as gloves, lab coat, goggles etc. All lab work is done at least in pairs. The labs are equipped with a controlled ventilating system, chemical fume hood, bio-safety cabinet, first aid kit, fire extinguisher, eye washer etc. Nevertheless, no personal items are allowed in the lab area. Also, eating and/or drinking are prohibited inside of the lab.

Bio Safety

All the microorganisms we are using are nonpathogenic.

Bacillus subtilis PY79 & 168

The bacterial strains are considered GRAS (Generally Considered As Safe). If somehow the inserted plasmid will be passed on to another sporulating bacteria (fully or partially), besides obtaining ampicillin resistance, the worst-case scenario is that the bacteria will have the ability to present ACE2 (Angiotensin-converting-enzyme 2) or “Sybodies” proteins on its exterior spore coat, which should not cause any harm.

Human HEK-293

When working with mammalian cells, extra safety measures were taken (such as working in a biological hood located inside our designated tissue culture room). In a worst-case scenario exposure, an infection might occur. Infections in immunocompetent individuals are generally localized and therefore do not pose a life-threatening risk. Generalized infections can occur in immunocompromised individuals. The symptoms of such infections include fever, rhinitis, pharyngitis, tonsillitis, cough and conjunctivitis^[1].

Saccharomyces Cerevisiae W303

The used yeast will not contain any yeast antibiotic resistance granting plasmid (we're using a different selection process: they are all mutagenic and lack an amino-acid related gene) so they can't transmit any harmful abilities.

Either way, all waste from working with microorganisms was autoclaved before being thrown away.

Product Safety

As a team, we aim our product to meet the FDA regulations regarding skin protectant drug products for over-the-counter human use. The main product safety issues that we tackle to achieve our goal derive from four hurdles.

Nickel

Our microgel beads contain Ni+ intended to attach the His-tag tail we fused to our proteins. Nickel is a well-known carcinogen by inhalation (gaseous form) and its metal form is suspected as one too^[2]. By skin contact Nickel can also cause eczema or pompholyx (both are skin allergy types). In a review published by the FDA concerning Nickel usage in internal implants, it is stated that for topical application (on skin), tests such as the Buehler Method and Human repeat inhalation patch test (HRIPT) can be used to assess skin sensitization in humans for different materials^[3]. Due to the fact we have not found any reports relating to skin sensitization in bound Nickel topical use, we are relying on the fact that the Nickel in our product is strongly bound^[4]. Also, if such a product will ever be planned to enter the market, clinical trials might be needed to determine the safety of the product (possible tests were described above). The trials should also include meticulous testing to make sure there is no Nickel "leakage".

Bacillus subtilis PY79 spores

Bacillus subtilis is GRAS. The FDA recognizes the Bacillus subtilis spores (strain SG118) as GRAS and allows use in food products^[5]. We therefore assume that the strain we use will also be approved by the FDA, especially because it is for topical application instead of food components. Vegetative Bacillus cells will be useless in the product, so germination needs to be avoided as much as possible. Therefore, we contacted two iGEM teams (Munich 2012^[6] & Freiburg 2016^[7]) that used the same non-germinating Bacillus subtilis strain in their project (developed by the Munich team) in hope to receive said strain (though the attempt was unsuccessful so far).

Pluronic F127

A hydrophilic non-ionic surfactant that is sometimes used in pharmaceuticals as a solubilizing and stabilizing agent. This substance is also known as Poloxamer 407 and there is a wide variety of FDA approved drugs containing it. The dose restriction (or amount threshold) is noted only for ophthalmic products^[8] (as far is we could find). In our product, the Pluronic F127 is the main ingredient of the hydrogel, which is the binding element for all our product's parts (hydrogel-based skin screen).

Misguidance

Our product is not a replacement for social distancing or masks. It is meant to be an additional aid and we created it while aiming to create a more appropriate and helpful solution rather than other sanitizers or gloves. If our product ever becomes commercially available, we will make sure that the message is clear to all users – You can use our protecting cream only as a replacement for gloves and constant hand sanitation, not anything else.

References

HEK 293 cell lines - Risk summary and guidelines for risk management. Biosafety. URL: https://biosafety.wsu.edu/hek-293-cell-lines/ Accessed tade: 18.09.2020
Of AR, Carcinogens H, Monographs I, Evaluation ONTHE, Risks OFC, Humans TO. And Dusts. Vol 100.; 2012. https://www.iarc.fr/.
US FDA. Biological Responses to Metal Implants.; 2019.
1. Mizrahi B, Irusta S, McKenna M, et al. Microgels for Efficient Protein Purification. Adv Mater. 2011;23(36):H258-H262. doi:10.1002/adma.201101258
GRAS Notice Inventory. U.S Food & Drug administration. URL: https://www.fda.gov/food/generally-recognized-safe-gras/gras-notice-inventory. Accessed tade: 18.09.2020
2012 M. How do Sporulation & Germination Work? https://2012.igem.org/Team:LMU-Munich/Germination_Stop/Knockout.
Freiburg 2016. Knock-Out. https://2016.igem.org/Team:Freiburg/Knockouts.
FDA alerts health care professionals of adverse events associated with Guardian’s compounded triamcinolone and moxifloxacin product for intravitreal injection. U.S Food & Drug administration. URL: https://www.fda.gov/drugs/drug-safety-and-availability/fda-alerts-health-care-professionals-adverse-events-associated-guardians-compounded-triamcinolone. Accessed tade: 18.09.2020

[r1] HEK 293 cell lines - Risk summary and guidelines for risk management. Biosafety. URL: https://biosafety.wsu.edu/hek-293-cell-lines/ Accessed tade: 18.09.2020

[r2] Of AR, Carcinogens H, Monographs I, Evaluation ONTHE, Risks OFC, Humans TO. And Dusts. Vol 100.; 2012. https://www.iarc.fr/.

[r3] US FDA. Biological Responses to Metal Implants.; 2019.

[r4] 1. Mizrahi B, Irusta S, McKenna M, et al. Microgels for Efficient Protein Purification. Adv Mater. 2011;23(36):H258-H262. doi:10.1002/adma.201101258

[r5] GRAS Notice Inventory. U.S Food & Drug administration. URL: https://www.fda.gov/food/generally-recognized-safe-gras/gras-notice-inventory. Accessed tade: 18.09.2020

[r6] 2012 M. How do Sporulation & Germination Work? https://2012.igem.org/Team:LMU-Munich/Germination_Stop/Knockout.

[r7] Freiburg 2016. Knock-Out. https://2016.igem.org/Team:Freiburg/Knockouts.

[r8] FDA alerts health care professionals of adverse events associated with Guardian’s compounded triamcinolone and moxifloxacin product for intravitreal injection. U.S Food & Drug administration. URL: https://www.fda.gov/drugs/drug-safety-and-availability/fda-alerts-health-care-professionals-adverse-events-associated-guardians-compounded-triamcinolone. Accessed tade: 18.09.2020

Team:Technion-Israel/Safety