Cell Line Engineering

Directing glycobiotechnology for industrial applications

The Cell Line Engineering group focuses on being at the frontier of Glycobiotechnology and its industrial applications. We engineer novel CHO cells lines, signal peptides, selection markers, and systems to enhance productivity and efficacy of recombinant therapeutics while boosting cell line stability and productivity. Through our work, we are working towards generating significant cost-effective and bioprocessing advantages for biosimilars and biotherapeutics.

Focus Areas

Genome editing of mammalian cell lines
Glycoengineering of CHO cells
Attenuated GS as selection markers in CHO cells for enhanced biologics production

CHO CELL GENOME EDITING AND EXPRESSION MODIFICATION

Using ZFNs, TALENS, CRISPR/Cas9 editing and RNA interference for boosting CHO lifespan, higher viable cell densities, productivity, and efficacy.

FUNCTIONAL STUDIES OF GLYCOSYLATION GENES

For streamlined glycoengineering efforts in key mammalian cell lines (e.g. CHO, HEK293, BHK, 3T3, NS0, Cos-7 etc.).

GLYCOENGINEERED CHO MUTANT CELL PANEL

Production of novel food associated oligosaccharide.

PRODUCTION OF ANTIBODY AND RECOMBINANT PROTEIN THERAPEUTICS WITH HOMOGENOUS N-GLYCANS

Enhance drug efficacy, suitable for chemical modifications such as generating ADCs.

Our deep experiences in glycoengineering facilitates success in protein production, particularly in view of protein glycosylation’s complexity, where more than 600 genes are estimated to be involved, compounded further by bioprocessing variability and industrial practicality.

GLYCOENGINEERING FOR PRODUCTION OF ANTIBODY AND RECOMBINANT PROTEIN THERAPEUTICS WITH HOMOGENOUS N-GLYCANS

CHO-gmt3 produced fucose-free antibody rituximab which killed cancer cells much more efficiently than commercial rituximab (MabThera)

GS-CHO system for biologics production

Comparison of attenuated GS R324C with GS wild type (WT) and with MSX for the stable production of mAb. (A) Antibody titer measurement of independently transfected stable cell pools. Stability assessment of mAb production over 8 weeks(>60 generations) of single clones generated by (B) GS WT+MSX and (C) GS R324C selection.

Featured Publications

Pao-Chun Lin, Ren Liu, Krista Alvin, Shahreel Wahyu, Nicholas Murgolo, Jianxin Ye, Zhimei Du and Zhiwei Song (2021) Improving Antibody Production in Stably Transfected CHO Cells by CRISPR-Cas9-Mediated Inactivation of Genes Identified in a Large-Scale Screen with Chinese Hamster-Specific siRNAs. Biotechnology Journal 16(3):e2000267
Sandra Prior, Aghiles Boughetane, Bernard Fox, Melissa Hayman, Eleanor Atkinson, Peter Rigsby, Meenu Wadhwa and Simon Hufton, and participants in the collaborative study (2020) Report on a Collaborative Study for Proposed Candidate 1st International Standard for the biological activities of Trastuzumab. WHO/BS/2020.2401
Pao-Chun Lin, Kah Fai Chan, Irene A Kiess, Joselyn Tan, Wahyu Shahreel, Sze-Yue Wong and Zhiwei Song (2019) Attenuated glutamine synthetase as selection marker in CHO cells to efficiently isolate highly productive stable cells for production of antibodies and other biologics. mAbs 11(5):965-976
Kah Fai Chan, Wahyu Shahreel, Corrine Wan, Gavin Teo, Noor Hayati, Shi Jie Tay, Wen Han Tong, Yuansheng Yang, Pauline M Rudd, Peiqing Zhang and Zhiwei Song (2016) Inactivation of GDP-fucose transporter gene (Slc35c1) in CHO cells by ZFNs, TALENs and CRISPR-Cas9 for production of fucose-free antibodies. Biotechnology Journal 11(3):399-414
John S Y Goh, Peiqing Zhang, Kah Fai Chan, May May Lee, Sing Fee Lim and Zhiwei Song (2010) RCA-I-resistant CHO mutant cells contain dysfunctional GnT I and expression of GnT I in these mutants enhances sialylation of recombinant erythropoietin. Metabomic Engineering 12(4):360-368

Landmark Patent & IP:

Attenuated glutamine synthetase as a selection marker (2018)
Optimised heavy chain and light chain signal peptides for the production of recombinant antibody therapeutics (2014)
Method of producing recombinant proteins with mannose-terminated N-glycans (2011)
Chinese hamster ovary cell linesGnT I mutant CHO cell lines (2009)

Dr Song Zhiwei

song_zhiwei@bti.a-star.edu.sg
Senior Principal Scientist II

PhD in Biochemistry (1993) University of Michigan, Ann Arbor, USA

Research Focus / Interest

Glycosylation of recombinant protein drugs
Apoptosis in cultured mammalian cells
Developing cell lines to produce follow-on biologics such as therapeutic antibodies

Dr Wang Huashan

wang_huashan@bti.a-star.edu.sg
Senior Scientist II

PhD in Molecular genetics (2006), Yong Loo Lin School of Medicine, National University of Singapore, Singapore

Research Focus / Interest

Expression of anticancer antibodies in cell lines
Development of cell lines for the production of therapeutic antibodies

Dr Chan Kah Fai

chan_kah_fai@bti.a-star.edu.sg
Senior Scientist I

PhD in Biochemistry (2017), Yong Loo Lin School of Medicine, National University of Singapore

Research Focus / Interest

Genetic engineering of novel CHO cell lines for industrial manufacturing of biologics
Applications of BTI CHO glycosylation mutant cell lines for production of alternative biomolecules
Recombinant fusion proteins and antibodies for therapeutic application

Cell Line Engineering

Directing glycobiotechnology for industrial applications

Focus Areas

Our Capabilities

CHO CELL GENOME EDITING AND EXPRESSION MODIFICATION