title: Production of monoclonal antibodies for breast cancer by HB8696 hybridoma cells using novel perfusion system.
abstract: Perfusion culture using spinfilters have been used for the production of health-care products using mammalian cells culture. However, available spinfilters are either highly prone to clog and/or are disposable and hence affects product formation. To address these problems, a novel non-woven Bombyx mori silk screen based spinfilter module for clog-free extended perfusion culture of hybridoma cells has been designed. The module is versatile in nature and reusable, after autoclaving and replacement of used polymeric membrane. Its application for clog-free extended perfusion culture was demonstrated by comparative perfusion experiments of HB8696 cells with stainless-steel spinfilter. HB8696 cells produce monoclonal antibodies (MAbs) 520C9 active against breast cancer oncoprotein. Silk spinfilter was found to be less prone to clog with cells and debris owing to its negatively charged hydrophobic screen compared to the positively charged hydrophilic stainless-steel spinfilter. Therefore, it provides extended cell growth phase and production phase of up to 56 h and 40 h respectively and 57.4% increase in MAb productivity compared to the stainless-steel spinfilter. The effect of different perfusion rates on MAb production was studied and an optimal MAb productivity of 1.6 g L(-1) day(-1) was achieved.
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
authors.forename: Shweta
authors.initials: S
authors.lastname: Kamthan
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology, Hauz Khas, New Delhi 110016, India. Electronic address: jgomes@bioschool.iitd.ac.in.
authors.forename: James
authors.initials: J
authors.lastname: Gomes
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India. Electronic address: pkrc@dbeb.iitd.ac.in.
authors.forename: Pradip K
authors.initials: PK
authors.lastname: Roychoudhury
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis*; D001709:Biotechnology; D001943:Breast Neoplasms / Q000276:immunology*; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D005260:Female; D005374:Filtration / Q000295:instrumentation; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D008855:Microscopy, Electron, Scanning; D010477:Perfusion; D016062:Porosity; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology; D047011:Silk / Q000648:ultrastructure; D013193:Stainless Steel
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D007074:Immunoglobulin G; D018719:Erb-b2 Receptor Tyrosine Kinases; D047011:Silk; D013193:Stainless Steel
keywords: Breast cancer; Extended perfusion operation; Filter clogging; Monoclonal antibody production; Silk spinfilter; Stainless-steel spinfilter