Lab safety
ABOA 2025 follows in the footsteps of the ABOA teams before us regarding safety measures. We are emulating the same clear division of topics for our safety text as it is easily legible, which makes it easier for future teams to learn from and build upon our team’s work. This year, as our project required working with human blood, it was especially important for our team to have very clear safety measures. Specifically, waste management, protecting our lab members from infection risks, and data safety concerning the blood samples were high on our priority list. Otherwise, we worked in the same laboratory as ABOA 2024 and have referenced their safety text when writing ours, updating it with the necessary extra information. We also follow all the same faculty guidelines from the Department of Molecular Plant Biology for personal safety, waste management, etc. Naturally, Finnish and EU laws concerning GMOs were taken into account in all our work. We also received additional guidance for working with human blood from Professor Kaisa Linderborg. For other lab-related risks, we had Senior Laboratory Technician Laura Kovesjoki and Laboratory Supervisor Mika Keränen by our side to give us expert advice whenever necessary.
Specialized safety measures for VeriFied
Working with human blood samples we needed specific rules for waste management, ethical handling of
data and biosafety, including infection risks. For biosafety, our lab is primarily labeled as BSL-1 so
extra precautions were taken in order to guarantee protection of BSL-2 or higher for everyone working in
the lab. We had a fume hood specifically reserved for working with blood samples, which was clearly
marked, as were all cabinets and drawers used for storing them.
Pictures of specified areas of the lab reserved for working with and storing biohazardous materials.
The sash of the fume hood was kept below shoulder height at all times and tubes containing the samples
were never opened outside this hood. In addition, blood samples were never handled by people without
Hepatitis-B vaccinations and appropriate PPE, including nitrile gloves over cotton gloves, specific
lab coats and protective eyewear. All personnel handling blood samples were trained and acted according
to safety guidelines set by the Department of Life Technologies Molecular Plant Biology. As an
additional safety measure, a 1mL blood sample was stored in case of an accidental exposure to blood,
so the hospital can run an array of tests on it. These measures were all reviewed and approved by
the Vice Head of the Department of Life Sciences in Turku, Kaisa Linderborg.
Blood samples were supplied to us by the Biotechnology department, where the blood samples are
collected routinely from volunteers - who have given written informed consent - in a completely
anonymous process. We did not sequence DNA from these samples, as it was not required for our project
and also to retain anonymity for the donors. The blood samples were only stored for the duration of
the lab work and were disposed of according to biohazardous waste management guidelines after all
blood-related work was done.
During our project we didn’t use any needles. The blood was stored in single-use plasticware,
samples being in the form of cotton swabs saturated with blood and stored in sealed microcentrifuge
tubes. All waste that came into any contact with blood was treated as biohazard waste and disposed
into special trash receptacles clearly marked with biohazard signs, in compliance with institutional
standards. Both solid and liquid waste were collected, weighed and recorded in a Biohazard Inventory
Form and then labeled. The waste was then incinerated in a designated cold storage facility.
A check-in form regarding human blood use was submitted to iGEM and approved before starting any lab work.
Personal safety
Before starting work in the lab, all lab personnel were required to complete an orientation and safety
training session. Laura Kovesjoki, the senior lab technician from the department of Molecular
Plant Biology, personally oversaw the sessions and signed binding forms with all attendees making sure
all necessary information was shared. No one is allowed to work in the labs without having attended
one of these orientation rounds.
Everyone in our lab team was familiar with lab etiquette through academic studies, reducing the risk
of “beginner mistakes”. Our lab was equipped with necessary safety features and we were shown where to
find emergency exits, showers, fire extinguishers, alarms and all other safety measures available in
the lab spaces.
Safety elements in our lab space; reused pictures of the same lab space from ABOA 2024.
All our protocols were drafted and checked before starting work in the lab. Our supervisors and PI
were consulted frequently to make sure everything was done in the lab as safely and efficiently as
possible.
As stated above, PPE and vaccines were especially important for team members handling blood samples,
but during handling of any hazardous chemicals nitrile gloves, lab coats and safety glasses were
required. Work involving hazardous chemicals was exclusively done in fume hoods. Long hair was always
to be tied neatly in a ponytail or bun, eating and drinking were strictly prohibited and washing hands
regularly was mandatory. To make sure to minimize the risk of further injury in the case that some
dangerous chemical got into someone’s eyes, contact lenses were prohibited. During possible exposure
of dangerous chemicals to the skin or eyes, the area should be immediately rinsed thoroughly.
All visitors were made aware of the safety protocols in the lab and acted accordingly.
Waste management
Waste management in our lab was carried out according to the system in place at PharmaCity with
additional practices set for biohazardous waste. All lab personnel were made aware of the appropriate
waste management protocols and adhered to them diligently. Any waste that had come into contact
with bacterial matter, including growth medium and pipette tips, were disposed of as detailed by
PharmaCity, following national GMO regulations. All waste in the lab was disposed of in an appropriate
manner, liquids were collected in clearly labeled containers, non-hazardous plastic waste and metals
recycled according to instructions available in the lab.
Handling of biohazardous samples and biohazard waste management in the lab.
AI and other concerns
AI was used at multiple points in our project, mostly to expedite brainstorming or help summarize large amounts of text. ChatGPT was the main AI tool used throughout the project. AI should always be used with caution and all sources checked thoroughly, as large language models often hallucinate sources or they may oversimplify information to the point of inaccuracy, sometimes even making confident false claims based on falsified or non-existent data. Keeping this in mind, our team utilized ChatGPT as a starting point, never a primary source of information, and all claims were comprehensively verified from additional sources. Large language models were also used to aid in coding for our wiki page, but we don’t foresee any issues, aside from ineffective code.
Chemicals used in our lab
There were plenty of chemicals used in our lab, for a multitude of different purposes, including some hazardous substances. For HIS-tag purification we used Co-NTA-resin, which contains Cobalt, which is classified as a heavy metal. DTT used for reducing HSA, imidazole that was utilized in IMAC purification and Ellman’s reagent that is an integral part of Ellman’s assay are all classified as carcinogens and mutagens, Ellman’s reagent also being classified as an acid/corrosive material and, along with imidazole, poisonous. Other acids and corrosive chemicals that were used are Page Blue (protein staining agent) used for SDS-PAGE, citric acid with which HSA was purified, HCl and KOH that were used in various buffers to adjust ph, sodium hypochlorite used as a cleaning agent and mercaptoethanol which was added to all SDS-PAGE samples. Out of these mercaptoethanol is also poisonous, in addition to ampicillin and kanamycin which are antibiotics that were used for selection and to avoid contamination with our bacteria. Ethanol which is standard for cleaning and sterilizing the lab is a highly flammable substance and the last hazardous chemical used in the lab.
Biosafety
Our chosen bacterial strain was BL21(DE3)pLysS E. coli, which is a specifically designed lab
strain, with specific growth requirements, to make it extremely unlikely to be able to compete with
other bacteria, if accidentally released. Additionally our engineered organism, or any part of it,
will not be part of the final test, as it is important for it to be completely cell-free to avoid
contamination on the crime scene. The organisms are used to express proteins (nanobody-NanoLuc fusions)
that will be purified and then utilized in the on-site test. Based on research and discussions with
experts from the Biotechnology department, the expressed proteins produced in the engineered strain
are unlikely to cause any environmental or physiological damage, as well as not providing any selective
advantage to the microbe.
The future of VeriFied
As a finalized product, VeriFied will be a cell-free on-site rapid test used to
determine bloodstain aging at a crime scene. The test would only be released to be used by certified
personnel. This would include crime scene investigators, police officers, or other on-scene professionals
able to use the test. The aim of VeriFied is to require minimal training and be easy
to use, so bloodstain age could be determined possibly even before crime scene investigators arrive
on-scene.
Using VeriFied should be simple: a sample is to be gathered from a dried bloodstain
using a cotton swab moistened with saline or sterile, cell-free water. Then the sample can be
extracted into a solution of substrate and buffer, which is added into test tubes (control and test)
containing dried proteins, and finally after a short incubation period their bioluminescence can be
measured with a portable luminometer, which will indicate age of the bloodstain according to signal
strength.
Our theoretical end goal for VeriFied is a totally usable new-generation tool for
crime scene investigation: a homogeneous enzyme complementation assay including purified nanobody-NanoLuc
fusion proteins expressed in genetically modified E. coli BL2(DE3)pLysS. All tests have been
performed only in our laboratory and no outside use has been tested.