Protocol

Glycosylation Engineering of Biopharmaceuticals

Volume 988 of the series Methods in Molecular Biology pp 31-43

Date:

Glycoengineered Pichia-Based Expression of Monoclonal Antibodies

  • Dongxing ZhaAffiliated withGlycoFi Inc., Merck and Co

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Currently, mammalian cells are the most commonly used hosts for the production of therapeutic monoclonal antibodies (mAbs). These hosts not only secrete mAbs with properly assembled two heavy and two light chains but also deliver mAbs with a glycosylation profile that is compatible with administration into humans. GlycoFi, a wholly owned subsidiary of Merck & Co., Inc., humanized the Pichia glycosylation pathway which allows it to express glycoproteins with a human-like glycan profile. This offers an alternative mAb production platform similar to mammalian hosts and in some cases it even provides more homogenous product and better efficacy, such as enhanced effector function. This chapter describes a protocol for using glycoengineered Pichia to produce full-length mAbs. It covers a broad spectrum of mAb expression technologies in yeast including expression vector construction, yeast transformation, high-throughput strain selection to fermentation, and antibody purification.

Key words

Glycoengineered Pichia Monoclonal antibody Glycosylation profile High-throughput screening FACS Fermentation and purification