Influence of glycosylation pattern on the molecular properties of monoclonal antibodies.

Abstract:

:Glycosylation is an important post-translational modification during protein production in eukaryotic cells, and it is essential for protein structure, stability, half-life, and biological functions. In this study, we produced various monoclonal antibody (mAb) glycoforms from Chinese hamster ovary (CHO) cells, including the natively glycosylated antibody, the enriched G0 form, the deglycosylated form, the afucosylated form, and the high mannose form, and we compared their intrinsic properties, side-by-side, through biophysical and biochemical approaches. Spectroscopic analysis indicates no measureable secondary or tertiary structural changes after in vitro or in vivo modification of the glycosylation pattern. Thermal unfolding experiments show that the high mannose and deglycosylated forms have reduced thermal stability of the CH2 domain compared with the other tested glycoforms. We also observed that the individual domain's thermal stability could be pH dependent. Proteolysis analysis indicates that glycosylation plays an important role in stabilizing mAbs against proteases. The stability of antibody glycoforms at the storage condition (2-8 °C) and at accelerated conditions (30 and 40 °C) was evaluated, and the results indicate that glycosylation patterns do not substantially affect the storage stability of the antibody we studied.

journal_name

MAbs

journal_title

mAbs

authors

Zheng K,Yarmarkovich M,Bantog C,Bayer R,Patapoff TW

doi

10.4161/mabs.28588

subject

Has Abstract

pub_date

2014-05-01 00:00:00

pages

649-58

issue

3

eissn

1942-0862

issn

1942-0870

pii

28588

journal_volume

6

pub_type

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