Affinity maturation of TCR-like antibodies using phage display guided by structural modeling

Author:

Frick Rahel123ORCID,Høydahl Lene S124ORCID,Hodnebrug Ina12,Vik Erik S5,Dalhus Bjørn6789,Sollid Ludvig M14,Gray Jeffrey J101112,Sandlie Inger12,Løset Geir Åge125ORCID

Affiliation:

1. Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital , Sognsvannsveien 20, 0372 Oslo, Norway

2. Centre for Immune Regulation and Department of Biosciences, University of Oslo , Blindernveien 31, 0371 Oslo, Norway

3. Department of Chemical and Biomolecular Engineering, Johns Hopkins University , 3400 N. Charles Street, Baltimore, MD 21218, USA

4. KG Jebsen Coeliac Disease Research Centre, University of Oslo , Sognsvannsveien 20, 0372 Oslo, Norway

5. Nextera AS , Gaustadalléen 21, 0349 Oslo, Norway

6. Department for Medical Biochemistry , Institute for Clinical Medicine, , Sognsvannsveien 20, 0372 Oslo, Norway

7. University of Oslo , Institute for Clinical Medicine, , Sognsvannsveien 20, 0372 Oslo, Norway

8. Department for Microbiology , Clinic for Laboratory Medicine, , Sognsvannsveien 20, 0372 Oslo, Norway

9. Oslo University Hospital , Clinic for Laboratory Medicine, , Sognsvannsveien 20, 0372 Oslo, Norway

10. Program in Molecular Biophysics, Johns Hopkins University , 3400 N. Charles Street, Baltimore, MD 21218, USA

11. Department of Chemical and Biomolecular Engineering and Institute of NanoBioTechnology, Johns Hopkins University , 3400 N. Charles Street, Baltimore, MD 21218, USA

12. Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins School of Medicine , 733 N Broadway, Baltimore, MD 21205, USA

Abstract

Abstract TCR-like antibodies represent a unique type of engineered antibodies with specificity toward pHLA, a ligand normally restricted to the sensitive recognition by T cells. Here, we report a phage display-based sequential development path of such antibodies. The strategy goes from initial lead identification through in silico informed CDR engineering in combination with framework engineering for affinity and thermostability optimization, respectively. The strategy allowed the identification of HLA-DQ2.5 gluten peptide-specific TCR-like antibodies with low picomolar affinity. Our method outlines an efficient and general method for development of this promising class of antibodies, which should facilitate their utility including translation to human therapy.

Funder

National Institutes of Health

Stiftelsen Kristian Gerhard Jebsen

South-Eastern Norway Regional Health Authority

Publisher

Oxford University Press (OUP)

Subject

Molecular Biology,Biochemistry,Bioengineering,Biotechnology

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