Production of monoclonal antibodies by glycoengineered Pichia pastoris.
The growing antibody market and the pressure to improve productivity as well as reduce cost of production have fueled the development of alternative expression systems. The therapeutic function of many antibodies is influenced by N-linked glycosylation, which is affected by a combination of the expression host and culture conditions. This paper reports the generation of a glycoengineered Pichia pastoris strain capable of producing more than 1 g l(-1) of a functional monoclonal antibody in a robust, scalable and portable cultivation process with uniform N-linked glycans of the type Man(5)GlcNAc(2). N-linked glycan uniformity and volumetric productivity have been maintained across a range of cultivation process conditions including pH (5.5-7.5), temperature (16-24 degrees C), dissolved oxygen concentration (0.85-3.40 mg l(-1)) and specific methanol feed rate (9-19 mg g(-1) h(-1)) as well as across different cultivation scales (0.5, 3.0, 15 and 40 l). Compared to a marketed CHO-produced therapeutic antibody, the glycoengineered yeast-produced antibody has similar motilities on SDS-PAGE, comparable size exclusion chromatograms (SEC) and antigen binding affinities. This paper provides proof of concept that glycoengineered yeast can be used to produce functional full-length monoclonal antibodies at commercially viable productivities.
Potgieter
Thomas I
TI
GlycoFi Inc. (A wholly-owned subsidiary of Merck & Co Inc.), 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA. Thomas Potgieter@Merck.com
Cukan
Michael
M
Drummond
James E
JE
Houston-Cummings
Nga Rewa
NR
Jiang
Youwei
Y
Li
Fang
F
Lynaugh
Heather
H
Mallem
Muralidhar
M
McKelvey
Troy W
TW
Mitchell
Teresa
T
Nylen
Adam
A
Rittenhour
Alissa
A
Stadheim
Terrance A
TA
Zha
Dongxing
D
d'Anjou
Marc
M
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D019149:Bioreactors; D002478:Cells, Cultured; D005818:Genetic Engineering; D042822:Genomic Instability; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000432:Methanol / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D013696:Temperature
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010100:Oxygen; D000432:Methanol