Presentation Title

Identification of Highly Selective MMP-14 Inhibitory Fabs by Deep Sequencing

Start Date

November 2016

End Date

November 2016

Location

HUB 379

Type of Presentation

Oral Talk

Abstract

Matrix metalloproteases (MMPs) are of rising interest in the treatment of cancer due to their roles in tumor growth and invasion. Current enrichment methods for monoclonal antibodies inhibiting specific MMPs involve phage panning and ELISA screening, which are usually associated with suboptimal growth conditions and insufficient expression levels of antibody clones. As the result, a significant proportion of potentially inhibitory clones might not be identified using classical screening procedures. Next Generation Sequencing (NGS) allows the rapid analysis of hundreds of millions of individual antibody clones in parallel. By monitoring the enrichment of clones along several rounds of phage panning against MMP-14 by using a MiSeq sequencer, we identified 17 new inhibitory clones not previously found using monoclonal phage ELISA. Particularly, Fab R2C7 exhibited an inhibition potency of 100 nM with an excellent selectivity to MMP-14 over MMP-9. Inhibition kinetics and epitope mapping analysis suggested that R2C7 is a competitive inhibitor and directly binds to the vicinity of MMP-14 catalytic site. In addition, several highly selective Fabs with inhibition IC50 values at 10-200 nM were also isolated. This study demonstrated that, as a supplementary means to ELISA, deep sequencing is very powerful to facilitate the systematic discovery of antibodies with protease inhibitory functions.

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Nov 12th, 10:00 AM Nov 12th, 10:15 AM

Identification of Highly Selective MMP-14 Inhibitory Fabs by Deep Sequencing

HUB 379

Matrix metalloproteases (MMPs) are of rising interest in the treatment of cancer due to their roles in tumor growth and invasion. Current enrichment methods for monoclonal antibodies inhibiting specific MMPs involve phage panning and ELISA screening, which are usually associated with suboptimal growth conditions and insufficient expression levels of antibody clones. As the result, a significant proportion of potentially inhibitory clones might not be identified using classical screening procedures. Next Generation Sequencing (NGS) allows the rapid analysis of hundreds of millions of individual antibody clones in parallel. By monitoring the enrichment of clones along several rounds of phage panning against MMP-14 by using a MiSeq sequencer, we identified 17 new inhibitory clones not previously found using monoclonal phage ELISA. Particularly, Fab R2C7 exhibited an inhibition potency of 100 nM with an excellent selectivity to MMP-14 over MMP-9. Inhibition kinetics and epitope mapping analysis suggested that R2C7 is a competitive inhibitor and directly binds to the vicinity of MMP-14 catalytic site. In addition, several highly selective Fabs with inhibition IC50 values at 10-200 nM were also isolated. This study demonstrated that, as a supplementary means to ELISA, deep sequencing is very powerful to facilitate the systematic discovery of antibodies with protease inhibitory functions.