7
From 2011.igem.org
Frank Machin
So the method for the anti-venom generation begins as follows:
1:
Take the venom proteins and bind biotin to either the C or N terminus so that it is able to be fixed to a large ferro-magnetic beads that are coated in streptavidin. Streptavidin has an extremely high affinity for biotin and the beads can be picked up with a neodynium magnet - so the bacteria with the 'antibody' on their surface that is able to bind to the venom proteins will be attached to the bead.
2:
The bacteria will have a plasmid that encodes a gene for a single-chain variable fragment, a fusion protein of the variable regions of the heavy and light chains of an immunoglobulin. This will be displayed upon the surface of the bacterium and will be on a plasmid that has a very mutagenic effect as described by Rebekka.
Rebekka Bauer
novel in vivo mutagenesis mechanism:
-construct target genomic DNA with really strong promoter with retrovirus recognition sequence (R U5 PBS etc to ensure that only this is reverse transcribed)
-transcribe into RNA using a crap RNA pol that introduces mutations
-reverse transcribe into DNA using an overexpressed reverse transcriptase (overexpressed to ensure that this happens before DNA is degraded). The RT should be coupled to an inducible promoter to ensure that mutagenesis only takes place in bursts.
-insert into genome via homologous recombination using an equivalent of infusion enzyme (maybe RecA?)
Atipat Patharagulpong
Contacts of people from RCA and LSE
RCA
- Pei-Ying Lin : peiying.lin@networkrca.ac.uk
- Koby : yaacov.barhad@network.rca.ac.uk
- CJ : charlotte@artforeating.com
LSE
- Alex Hamilton : r.a.hamilton@LSE.ac.uk
- Claire MArris : c.marris@lse.ac.uk
- Stephan Guttinges : s.m.guettinges@lse.ac.uk
Talk from LSE
Things to consider in human practise
Safety, Security, IP-intellectual property, ethical, biohacking, global fairness
For Anti-venom project
Safety : handling toxin in the lab, storage and transportation of toxins
Security : produce specificity problem, antivenom target for particular person
Patent : only the process obtaining product(antibody) but not the product itself, patent cost a lot of money