Diversity in host clone performance within a Chinese hamster ovary cell line.
Much effort has been expended to improve the capabilities of individual Chinese hamster ovary (CHO) host cell lines to synthesize recombinant therapeutic proteins (rPs). However, given the increasing variety in rP molecular types and formats it may be advantageous to employ a toolbox of CHO host cell lines in biomanufacturing. Such a toolbox would contain a panel of hosts with specific capabilities to synthesize certain molecular types at high volumetric concentrations and with the correct product quality (PQ). In this work, we examine a panel of clonally derived host cell lines isolated from CHOK1SV for the ability to manufacture two model proteins, an IgG4 monoclonal antibody (Mab) and an Fc-fusion protein (etanercept). We show that these host cell lines vary in their relative ability to synthesize these proteins in transient and stable pool production format. Furthermore, we examined the PQ attributes of the stable pool-produced Mab and etanercept (by N-glycan ultra performance liquid chromatography (UPLC) and liquid chromatography - tandem mass spectrometry (LC-MS/MS), respectively), and uncovered substantial variation between the host cell lines in Mab N-glycan micro-heterogeneity and etanercept N and O-linked macro-heterogeneity. To further investigate the capabilities of these hosts to act as cell factories, we examined the glycosylation pathway gene expression profiles as well as the levels of endoplasmic reticulum (ER) and mitochondria in the untransfected hosts. We uncovered a moderate correlation between ER mass and the volumetric product concentration in transient and stable pool Mab production. This work demonstrates the utility of leveraging diversity within the CHOK1SV pool to identify new host cell lines with different performance characteristics.
Lonza Biologics Plc, New Expression Technologies Group, Cambridge, CB21 6GS, U.K.
Peter M
PM
O'Callaghan
Lonza Biologics Plc, New Expression Technologies Group, Cambridge, CB21 6GS, U.K.
Maud E
ME
Berthelot
Lonza Biologics Plc, New Expression Technologies Group, Cambridge, CB21 6GS, U.K.
Robert J
RJ
Young
Protein Chemistry, Lonza Biologics Plc, Slough, SL1 4DX, U.K.
James W A
JW
Graham
Process Development Sciences, Lonza Biologics Plc, Slough, SL1 4DX, U.K.
Andrew J
AJ
Racher
Process Analytics, Lonza Biologics Plc, Slough, SL1 4DX, U.K.
Dulce
D
Aldana
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000494:pharmacology; D016466:CHO Cells; D002853:Chromatography, Liquid; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D006031:Glycosylation; D007074:Immunoglobulin G / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011134:Polysaccharides; D012333:RNA, Messenger; D011994:Recombinant Proteins
Chinese hamster ovary cell line; biosimilar manufacturing; cell line variation; recombinant protein