Generation and evolution of human antibody fragments - The CDR shuffling approach

Sammanfattning: Antibodies are important proteins participating in a number of reactions to protect us from harmful diseases. Apart from this important function, they are also useful in different technical and medical applications. This, however, relies on our possibility to produce antibodies in vitro. In the first part of my thesis a new technology called CDR shuffling was developed to create diverse and highly functional antibody libraries. Specific antibody fragments were isolated from such CDR shuffling antibody libraries by phage display selections. When isolating antibody fragments from antibody libraries, the retrieved clones may not have the desired reactivity characteristics. It may be necessary to further in vitro evolve the clones to achieve the desired kinetic or specificity parameters. In the second part of my thesis, the CDR shuffling technology was used to further evolve initially isolated antibody fragments. By this method, I was able to define a central core structure important for generating a mucin-1 specificity. By keeping this central core and introducing variability in other parts of the molecules, I further improved the kinetic parameters of the isolated clones. In conclusion, this thesis presents a novel technology suitable for the generation and evolution of human antibody fragments.