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J Biol Chem. 2020 Jan 17. pii: jbc.RA119.011712. doi: 10.1074/jbc.RA119.011712. [Epub ahead of print]

Selective inhibition of matrix metalloproteinase 10 (MMP10) with a single-domain antibody.

Author information

1
SBP Medical Discovery Institute, United States.
2
Sanford Burnham Prebys Medical Discovery Institute, United States.

Abstract

Since their discovery, the matrix metalloproteinase (MMP) family of proteases have been considered as therapeutic targets in numerous diseases and disorders. Unfortunately, clinical trials with MMP inhibitors have failed to yield any clinical benefits of these inhibitors. These failures were largely due to a lack of MMP-selective agents; accordingly, it has become important to identify a platform with which high selectivity can be achieved. To this end, we propose using MMP-targeting antibodies that can achieve high specificity in interactions with their targets. Using a scaffold of single-domain antibodies, here we raised a panel of MMP10-selective antibodies through immunization of llamas, a member of the camelid family, whose members generate conventional heavy/light chain antibodies and also smaller antibodies lacking light-chain and CH1 domains. We report the generation of a highly selective and tightly binding MMP10 inhibitor (Ki < 2 nM). Using bio-layer interferometry-based binding assays, we found that this antibody interacts with the MMP10 active site.  Activity assays demonstrated that the antibody selectively inhibits MMP10 over its closest relative MMP3. The ability of a single-domain antibody to discriminate between the most conserved MMP pair via an active site-directed mechanism of inhibition reported here supports the potential of this antibody as a broadly applicable scaffold for the development of selective, tightly binding MMP inhibitors.

KEYWORDS:

antibody; antibody engineering; metalloprotease; protease inhibitor; single-domain antibody (sdAb,nanobody)

PMID:
31953328
DOI:
10.1074/jbc.RA119.011712
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