NISTmAb, Humanized IgG1κ Monoclonal Antibody

This reference material (RM) is intended primarily for use in evaluating the performance of methods for determining physicochemical and biophysical attributes of monoclonal antibodies.

It also provides a representative test molecule for development of novel technologies for therapeutic protein characterization.

This RM can be used for a variety of purposes that may include system suitability tests, establishing method or instrument performance and variability, comparing changing analytical test methods, and assisting in method qualification.

A unit of RM 8671 consists of one internal-threaded polypropylene cryovial containing 800 µL of 10 mg/mL NIST IgG1κ monoclonal antibody (NISTmAb) in 12.5 mmol/L L-histidine, 12.5 mmol/L L-histidine HCl (pH 6.0). 

The NIST monoclonal antibody (NISTmAb) is a recombinant humanized IgG1κ expressed in murine suspension culture and has undergone biopharmaceutical industry standard upstream and downstream purification to remove process related impurities.

It is an ~150 kDa homodimer of two light chain and two heavy chain subunits linked through both inter- and intra-chain disulfide bonds.

The molecule has a high abundance of N terminal

pyroglutamination, C terminal lysine clipping, and glycosylation of the heavy chains. The protein also has low abundance post translational modifications including methionine oxidation, deamidation, and glycation.

These and other product quality attributes were extensively characterized in the ACS Symposium Series for the initial batch of interim material, used as the primary sample (PS) herein.