Molecular imprint of enzyme active site by camel nanobodies: rapid and efficient approach to produce abzymes with alliinase activity.

Abstract

Screening of inhibitory Ab1 antibodies is a critical step for producing catalytic antibodies in the anti-idiotypic approach. However, the incompatible surface of the active site of the enzyme and the antigen-binding site of heterotetrameric conventional antibodies become the limiting step. Because camelid-derived nanobodies possess the potential to preferentially bind to the active site of enzymes due to their small size and long CDR3, we have developed a novel approach to produce antibodies with alliinase activities by exploiting the molecular mimicry of camel nanobodies. By screening the camelid-derived variable region of the heavy chain cDNA phage display library with alliinase, we obtained an inhibitory nanobody VHHA4 that recognizes the active site. Further screening with VHHA4 from the same variable domain of the heavy chain of a heavy-chain antibody library led to a higher incidence of anti-idiotypic Ab2 abzymes with alliinase activities. One of the abzymes, VHHC10, showed the highest activity that can be inhibited by Ab1 VHHA4 and alliinase competitive inhibitor penicillamine and significantly suppressed the B16 tumor cell growth in the presence of alliin in vitro. The results highlight the feasibility of producing abzymes via anti-idiotypic nanobody approach.

• Authors: Jiang-Wei Li, Lijie Xia, Youhong Su, Hongchun Liu, Xueqing Xia, Qinxia Lu, Chunjin Yang, Kalbinur Reheman
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 287 Issue 17 Pg. 13713-21 (Apr 20 2012) ISSN: 1083-351X [Electronic] United States
• PMID: 22374998 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• Antibodies, Anti-Idiotypic
• Enzyme Inhibitors
• Enzymes
• Peptide Library
• Carbon-Sulfur Lyases
• alliin lyase
• Penicillamine

Topics

• Animals
• Antibodies (chemistry)
• Antibodies, Anti-Idiotypic (chemistry)
• Binding Sites
• Camelus
• Carbon-Sulfur Lyases (chemistry)
• Catalysis
• Catalytic Domain
• Enzyme Inhibitors (chemistry)
• Enzyme-Linked Immunosorbent Assay (methods)
• Enzymes (chemistry)
• Female
• Kinetics
• Molecular Sequence Data
• Nanotechnology (methods)
• Neoplasms (immunology, metabolism)
• Penicillamine (chemistry)
• Peptide Library