Isolation of Antigen-Specific Unconventional Bovine Ultra-Long CDR3H Antibodies Using Cattle Immunization in Combination with Yeast Surface Display

Paul Arras; Jasmin Zimmermann; Britta Lipinski; Desislava Yanakieva; Daniel Klewinghaus; Simon Krah; Harald Kolmar; Lukas Pekar; Stefan Zielonka

doi:10.1007/978-1-0716-3279-6_8

Isolation of Antigen-Specific Unconventional Bovine Ultra-Long CDR3H Antibodies Using Cattle Immunization in Combination with Yeast Surface Display

Methods Mol Biol. 2023:2681:113-129. doi: 10.1007/978-1-0716-3279-6_8.

Authors

Paul Arras¹, Jasmin Zimmermann^{1

2}, Britta Lipinski^{1

2}, Desislava Yanakieva¹, Daniel Klewinghaus¹, Simon Krah¹, Harald Kolmar², Lukas Pekar¹, Stefan Zielonka^{3

4}

Affiliations

¹ Antibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, Germany.
² Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany.
³ Antibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, Germany. [email protected].
⁴ Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany. [email protected].

PMID: 37405646
DOI: 10.1007/978-1-0716-3279-6_8

Abstract

Cattle are known for their repertoire of antibodies harboring extremely long CDR3H regions that form extensive "knob on stalk" cysteine-rich structures. The compact knob domain allows for the recognition of epitopes potentially not accessible to classical antibodies. To effectively access the potential of bovine-derived antigen-specific ultra-long CDR3 antibodies, a straightforward and effective high-throughput method based on yeast surface display and fluorescence-activated cell sorting is described.

Keywords: Antibody display; Antibody engineering; Bovine ultra-long CDR3H antibodies; Stalk-knob; Yeast surface display.

MeSH terms

Animals
Antibodies*
Antigens
Cattle
Immunization
Saccharomyces cerevisiae* / genetics
Vaccination

Substances

Antibodies
Antigens