In vitro antibody affinity maturation using germinal centre organoids

This project will develop a rapid method to improve antibodies without the need to use an animal model of immunisation.

Antibodies are widely used in industrial processes, diagnostic tests and as medicines because they provide an incredibly precise way to target specific molecules.

However, developing antibodies within the bioscience sector is time consuming and expensive, often because the original antibody discovered almost always has to be optimised to make it fit for purpose.

Currently this optimisation involves either repeating the animal immunisations to identify relatives of the original antibody that have a better profile for development into a medicine/biological tool, or a lengthy and unphysiological process to mutate the antibody for improved performance.

Here, instead, we will use a new lab-based organoid system that mimics the normal environment of antibody optimisation within the body by using tonsil cells discarded after routine operations.

We will genetically engineer the tonsil cells using Crispr-cas9 to make the antibody that needs optimisation and then trigger the antibody optimisation process in the lab organoid.

The first objective is to genetically engineer the type of tonsil cells that normally make antibodies by removing their original antibody and replacing it with different antibodies in turn and showing that the organoid system continues to work after the genetic engineering.

The second objective is to establish the best laboratory conditions for optimising the antibody function in terms of how strongly they attach to the target molecule.

These objectives will be reached by using four well-studied antibodies against a target from a virus (HIV) that none of the donors providing tonsil tissues have experienced.

Therefore, we will be able to conclusively prove that it is the genetic insertion of the antibodies that is leading to improved antibody function in the organoid, rather than the organoid optimisation working on pre-existing antibodies from the donor.

Moreover, as the four antibodies and other antibodies in their family tree have been previously studied we can benchmark the progress in the organoid model against what has already happened in natural antibody maturation.

This project will establish the proof-of-concept for a new technology which could allow much more rapid (weeks rather than months) improvement of antibodies with consequently less environmental impact and without the experimental animals.

Antibodies improved by this method could be used to treat/detect cancer and other diseases in humans and livestock and as crucial components in diagnostic and industrial processes.