Phenotypic antibody discovery and mining of complex antibody libraries
Summary, in English
In this thesis, I establish a phenotypic antibody discovery platform and apply it on primary cancer cells from patients with chronic lymphocytic leukemia, CLL. Novel antibody-target combinations, with enhanced cytotoxicity, both in vitro and in vivo, compared to the standard of care were discovered. Furthermore, I showed that by applying various deep mining methods on the generated antibody pool, additional antibodies could be discovered. These rare antibodies bound new epitopes on the target cells, either on previously discovered, or on entirely new targets. In addition, I demonstrate through analysis by next generation sequencing, NGS, that the number of receptors on the cell surface has a major impact on antibody enrichment in the selection process. By following clonal enrichment with NGS, both abundant and rare antibodies were discovered. This allows the conventional immunochemical screening of a phage display antibody discovery process to be by-passed.
The discovered antibodies can be used in many different applications. For some of these applications, bispecific antibodies, designed to bind two different antigens, can have improved efficacy through e.g. increased specificity compared to conventional monospecific antibodies. However, many of the current bispecific formats, especially those similar to a conventional IgG, are difficult to construct. In this thesis, I create a new type of bispecific antibody, the Tetra-VH IgG. This antibody format contains four variable heavy (VH) domains, is tetravalent, and binds two antigens simultaneously on each Fab arm. The Tetra-VH IgGs potentially show enhanced binding and functional properties compared to conventional bivalent IgGs.
In summary, the technologies I describe in this thesis will be important in future antibody discovery efforts as they enable a broad repertoire of antibodies against novel targets, of both high and low receptor density, to be discovered and used for treatment of various diseases.
- Immunology in the medical area
- Biochemistry and Molecular Biology
- Antibody discovery
- Phage display
- Phenotypic screening
- Cell selection
- Deep mining
- Complex phage pools
- ISBN: 978-91-7895-349-3
- ISBN: 978-91-7895-348-6
24 januari 2020
Lecture hall MA2, Matteannexet, Sölvegatan 20, Faculty of Engineering LTH, Lund University, Lund.
- Gregory. Winter (Sir.)