Team:Potsdam Bioware/Project/At a Glance

At a Glance

Antibody Generation System

The main goal of our antibody generation system is to produce high affine antibody using an antibody module, a mutation module and a selection module to ensure that the cells which expressed a high affine antibody survive grow.
For the antibody module, we transiently and stable transfected CHO cells with two antibody constructs. The first one consists of a single chain against the epidermale growth factor receptor, a transmembrane region and a signal peptide. The latter ones ensure that the construct is presented on the cell surface. The second one contains a nanobody against GFP a transmembrane region and a signal peptide with the same function like above and also pseudo intron region to ensure the switch between soluble and membrane state of the antibody’s.
The mutation module consists of one key enzyme, the activation induced cytidine deaminase (AID). This enzyme is commonly used in mammalian immune systems to induced the hypermuation and thus antibody maturation in activated B-lymphocytes. We used the wildtype form and a modified variant of this enzyme with a nuclear localization sequence and without a nuclear export sequence for transfection in CHO cells to induce hypermutation. The transfected AID induce hypermutation in antibody transfected CHO cells and thus change the antibody binding regions stochastically.
To select CHO cells which produce a high affine antibody, we constructed a selection module. This module consists of viruses which show the corresponding antigen, for the nanobody GFP, on the surface by using a fusion protein. The virus has an antibiotic resistance cassette. By binding the high affine antibody with the surface presenting antigen the virus is able to infect the CHO cells effectively. Consequently, only the CHO cells survive which produce a high affine antibody mutated by the AID.

Collaboration

We are working together with the University Freiburg team to test their Transactivator-like (TAL) proteins by sending one of our modified AID enzymes for ligation to the TAL domain. Team Freiburg, on the other hand, plans to send back the TAL domain linked to the AID via glycine-serine linker. Afterwards, we aim to check the mutation rate of the specifically directed AID to our antibody sequence.

SocialBrick

The idea of SocialBrick is to divide all human practice acts into different parts: the SocialBricks. Here, the SocialBricks stands for every activity which aims to inform the people about the Synthetic Biology. We hope that the term SocialBrick will be accepted like the term Biobrick and will be integrated in a registry to show the society what every team has done for more elucidation.
For this year we focused on two SocialBricks: “Science meets Politics” and “Science meets People”. For the first one we interviewed politicians from the German parliament: the Bundestag and discussed about Synthetic Biology. For the second part we organized a survey to ask about people's knowledge of Synthetic Biology and for their opinion on this scientific field.
The main results of the human practice this year are that the citizens and the politicians see the great potential of Synthetic Biology but also the challenges of this new scientific field.