Physiologically relevant binding affinity quantification of monoclonal antibody PF-00547659 to mucosal addressin cell adhesion molecule for in vitro in vivo correlation.

ABSTRACT:

Background and purpose

A monoclonal antibody (PF-00547659) against mucosal addressin cell adhesion molecule (MAdCAM), expressed as both soluble (sMAdCAM) and trans-membrane (mMAdCAM) target forms, showed over 30-fold difference in antibody-target K_D between in vitro (Biacore) and clinically derived (K_D,in-vivo ) values. Back-scattering interferometry (BSI) was applied to acquire physiologically relevant K_D values which were used to establish in vitro and in vivo correlation (IVIVC).

Experimental approach

BSI was applied to obtain K_D values between PF-00547659 and recombinant human MAdCAM in buffer or CHO cells and endogenous MAdCAM in human serum or colon tissue. CHO cells and tissue were minimally processed to yield homogenate con

SUBMITTER: Wang M 

PROVIDER: S-EPMC5221447 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature