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Generation of highly selective monoclonal antibodies inhibiting a recalcitrant protease using decoy designs.

Authors
  • Lee, Ki Baek1
  • Dunn, Zachary S1, 2
  • Lopez, Tyler1, 3
  • Mustafa, Zahid1
  • Ge, Xin1
  • 1 Department of Chemical and Environmental Engineering, University of California Riverside, Riverside, California.
  • 2 Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, California.
  • 3 Element Biosciences, Inc., San Diego, California.
Type
Published Article
Journal
Biotechnology and Bioengineering
Publisher
Wiley (John Wiley & Sons)
Publication Date
Dec 01, 2020
Volume
117
Issue
12
Pages
3664–3676
Identifiers
DOI: 10.1002/bit.27519
PMID: 32716053
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Matrix metalloproteinase-12 (MMP-12), also known as macrophage elastase, is a potent inflammatory mediator and therefore an important pharmacological target. Clinical trial failures of broad-spectrum compound MMP inhibitors suggested that specificity is the key for a successful therapy. To provide the required selectivity, monoclonal antibody (mAb)-based inhibitors are on the rise. However, poor production of active recombinant human MMP-12 catalytic domain (cdMMP-12) presented a technical hurdle for its inhibitory mAb development. We hypothesized that this problem could be solved by designing an expression-optimized cdMMP-12 mutant without structural disruptions at its reaction cleft and surrounding area, and thus isolated active-site inhibitory mAbs could maintain their binding and inhibition functions toward wild-type MMP-12. We combined three advances in the field-PROSS algorithm for cdMMP-12 mutant design, convex paratope antibody library construction, and functional selection for inhibitory mAbs. As a result, isolated Fab inhibitors showed nanomolar affinity and potency toward cdMMP-12 with high selectivity and high proteolytic stability. Particularly, Fab LH11 targeted the reaction cleft of wild-type cdMMP-12 with 75 nM binding KD and 23 nM inhibition IC50 . We expect that our methods can promote the development of mAbs inhibiting important proteases, many of which are recalcitrant to functional production. © 2020 Wiley Periodicals LLC.

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