Antibody molecules are enormously important as therapeutic and diagnostic molecules. More recently, unique scaffolds like the VHH of camelids, or even non-antibody frameworks like knottins have become important in biomedicine. Antibody diversity and antigen binding in mammals is often restricted to the CDR loops of the immunoglobulin fold. In experiments challenging this paradigm, we have recently solved the crystal structures of two bovine antibodies containing ultralong CDR H3s (56 and 61 amino acids) and also deep sequenced the ultralong repertoire. Our data reveal that these CDR H3s form a very unusual architecture composed of a long ?-strand stalk which supports a disulfide rich knob that protrudes far from the immunoglobulin surface. Interestingly, the two different antibodies contain different patterns of disulfides, which result in different knob structures. Dee sequencing reveals extensive diversity in the ultralong CDR H3s where a multitude of different disulfides could potentially form within the knob. Thus, the bovine antibody system can produce an unprecedented repertoire of mega CDR H3s that may result in an impressive diversity of minifolds containing combinations of somatically generated disulfides. Thus, antibody diversity is located in a new minifold supported by the immunoglobulin domain. We will perform structural, functional, and engineering studies to investigate the properties of this new antibody class, as well as to lead the way to developing this unique structure into therapeutics.

Public Health Relevance

Understanding the structure and function of antibodies is enormously important in several medical fields. Antibody drugs are now a large therapeutic class, therefore detailed knowledge underlying antibody diversity, physical properties, affinity and engineering is critically important to drug development. Our research on the structure of a unique class of bovine antibodies with ultralong CDR3s will further develop our understanding of and ability to engineer the antibody molecule.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM105826-02
Application #
8930163
Study Section
Macromolecular Structure and Function B Study Section (MSFB)
Program Officer
Smith, Ward
Project Start
2014-09-30
Project End
2018-07-31
Budget Start
2015-08-01
Budget End
2016-07-31
Support Year
2
Fiscal Year
2015
Total Cost
$360,050
Indirect Cost
$170,050
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Haakenson, Jeremy K; Huang, Ruiqi; Smider, Vaughn V (2018) Diversity in the Cow Ultralong CDR H3 Antibody Repertoire. Front Immunol 9:1262
Sok, Devin; Le, Khoa M; Vadnais, Melissa et al. (2017) Rapid elicitation of broadly neutralizing antibodies to HIV by immunization in cows. Nature 548:108-111
Vadnais, Melissa L; Smider, Vaughn V (2016) Bos taurus ultralong CDR H3 antibodies. Curr Opin Struct Biol 38:62-7
Stanfield, Robyn L; Wilson, Ian A; Smider, Vaughn V (2016) Conservation and diversity in the ultralong third heavy-chain complementarity-determining region of bovine antibodies. Sci Immunol 1:
Muyldermans, Serge; Smider, Vaughn V (2016) Distinct antibody species: structural differences creating therapeutic opportunities. Curr Opin Immunol 40:7-13
de los Rios, Miguel; Criscitiello, Michael F; Smider, Vaughn V (2015) Structural and genetic diversity in antibody repertoires from diverse species. Curr Opin Struct Biol 33:27-41