Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 113, Issue 4, Pages 717-723Publisher
WILEY
DOI: 10.1002/bit.25840
Keywords
MMP; periplasmic expression; inhibitory antibody
Categories
Funding
- NSF [CBET 1453645]
- UC Cancer Research Coordinating Committee
- Directorate For Engineering [1453645] Funding Source: National Science Foundation
- Div Of Chem, Bioeng, Env, & Transp Sys [1453645] Funding Source: National Science Foundation
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Human matrix metalloproteinase (MMP)-14, a membrane-bound zinc endopeptidase, is one of the most important cancer targets because it plays central roles in tumor growth and invasion. Large amounts of active MMP-14 are required for cancer research and the development of chemical or biological MMP-14 inhibitors. Current methods of MMP-14 production through refolding and activation are labor-intensive, time-consuming, and often associated with low recovery rates, lot-to-lot variation and heterogeneous products. Here, we report direct production of the catalytic domain of MMP-14 in the periplasmic space of Escherichia coli. 0.5mg/L of functional MMP-14 was produced without tedious refolding or problematic activation process. MMP-14 prepared by simple periplasmic treatment can be readily utilized to evaluate the potencies of chemical and antibody-based inhibitors. Furthermore, co-expression of both MMP-14 and antibody Fab fragments in the periplasm facilitated inhibitory antibody screening by avoiding purification of MMP-14 or Fabs. We expect this MMP-14 expression strategy can expedite the development of therapeutic drugs targeting MMPs with biological significance. (c) 2015 Wiley Periodicals, Inc.
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