Rational design of a Kv1.3 channel-blocking antibody as a selective immunosuppressant.
A variable region fusion strategy was used to generate an immunosuppressive antibody based on a novel "stalk-knob" structural motif in the ultralong complementary-determining region (CDR) of a bovine antibody. The potent Kv1.3 channel inhibitory peptides Moka1-toxin and Vm24-toxin were grafted into different CDRs of the humanized antibodies BVK and Synagis (Syn) using both β-sheet and coiled-coil linkers. Structure-activity relationship efforts led to generation of the fusion protein Syn-Vm24-CDR3L, which demonstrated excellent selectivity and potency against effector human memory T cells (subnanomolar to picomolar EC50values). This fusion antibody also had significantly improved plasma half-life and serum stability in rodents compared with the parent Vm24 peptide. Finally, this fusion protein showed potent in vivo efficacy in the delayed type hypersensitivity in rats. These results illustrate the utility of antibody CDR fusions as a general and effective strategy to generate long-acting functional antibodies, and may lead to a selective immunosuppressive antibody for the treatment of autoimmune diseases.
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; California Institute for Biomedical Research, La Jolla, CA 92037.
Rongsheng E
RE
Wang
California Institute for Biomedical Research, La Jolla, CA 92037.
Ying
Y
Wang
California Institute for Biomedical Research, La Jolla, CA 92037.
Yuhan
Y
Zhang
California Institute for Biomedical Research, La Jolla, CA 92037.
Chase
C
Gabrelow
California Institute for Biomedical Research, La Jolla, CA 92037.
Yong
Y
Zhang
California Institute for Biomedical Research, La Jolla, CA 92037.
Victor
V
Chi
California Institute for Biomedical Research, La Jolla, CA 92037.
Qiangwei
Q
Fu
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037.
Xiaozhou
X
Luo
California Institute for Biomedical Research, La Jolla, CA 92037.
Danling
D
Wang
California Institute for Biomedical Research, La Jolla, CA 92037.
Sean
S
Joseph
California Institute for Biomedical Research, La Jolla, CA 92037.
Kristen
K
Johnson
California Institute for Biomedical Research, La Jolla, CA 92037.
Arnab K
AK
Chatterjee
California Institute for Biomedical Research, La Jolla, CA 92037.
Timothy M
TM
Wright
California Institute for Biomedical Research, La Jolla, CA 92037.
Vân T B
VT
Nguyen-Tran
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA 92037.
John
J
Teijaro
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA 92037.
Argyrios N
AN
Theofilopoulos
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; California Institute for Biomedical Research, La Jolla, CA 92037; schultz@scripps.edu fwang@calibr.org.
Peter G
PG
Schultz
California Institute for Biomedical Research, La Jolla, CA 92037; schultz@scripps.edu fwang@calibr.org.
Feng
F
Wang
D000595:Amino Acid Sequence; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology*; D016466:CHO Cells; D002417:Cattle; D022801:Complementarity Determining Regions / Q000737:chemistry; D006224:Cricetinae; D003412:Cricetulus; D015195:Drug Design; D057809:HEK293 Cells; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D051664:Kv1.3 Potassium Channel / Q000037:antagonists & inhibitors*; D008213:Lymphocyte Activation / Q000187:drug effects; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology
D019138:Antibodies, Blocking; D022801:Complementarity Determining Regions; D007166:Immunosuppressive Agents; D051664:Kv1.3 Potassium Channel; D011993:Recombinant Fusion Proteins
Kv1.3; antibody; autoimmune; immunosuppressive; protein engineering
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