title: Multiparameter optimization method and enhanced production of secreted recombinant single-chain variable fragment against the HIV-1 P17 protein from Escherichia coli by fed-batch fermentation.
abstract: The single-chain fragment variable (scFv) was used to produce a completely functional antigen-binding fragment in bacterial systems. The advancements in antibody engineering have simplified the method of producing Fv fragments and made it more efficient and generally relevant. In a previous study, the scFv anti HIV-1 P17 protein was produced by a batch production system, optimized by the sequential simplex optimization method. This study continued that work in order to enhance secreted scFv production by fed-batch cultivation, which supported high volumetric productivity and provided a large amount of scFvs for diagnostic and therapeutic research. The developments in cell culture media and process parameter settings were required to realize the maximum production of cells. This study investigated the combined optimization methods, Plackett-Burman design (PBD) and sequential simplex optimization, with the aim of optimize feed medium. Fed-batch cultivation with an optimal feeding rate was determined. The result demonstrated that a 20-mL/hr feeding rate of the optimized medium can increase cell growth, total protein production, and scFv anti-p17 activity by 4.43, 1.48, and 6.5 times more than batch cultivation, respectively. The combined optimization method demonstrated novel power tools for the optimization strategy of multiparameter experiments.
authors.affiliation: a Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.
authors.forename: Porntip
authors.initials: P
authors.lastname: Paopang
authors.affiliation: b Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.|c Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.
authors.forename: Watchara
authors.initials: W
authors.lastname: Kasinrerk
authors.affiliation: b Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.|c Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.
authors.forename: Chatchai
authors.initials: C
authors.lastname: Tayapiwatana
authors.affiliation: d Department of Biotechnology, Faculty of Agro-Industry , Chiang Mai University , Chiang Mai , Thailand.
authors.forename: Phisit
authors.initials: P
authors.lastname: Seesuriyachan
authors.affiliation: a Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences , Chiang Mai University , Chiang Mai , Thailand.
authors.forename: Bordin
authors.initials: B
authors.lastname: Butr-Indr
mesh_terms: D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D015488:HIV Antigens / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000276:immunology*
chemical_list: D015488:HIV Antigens; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D054301:gag Gene Products, Human Immunodeficiency Virus; C054858:p17 protein, Human Immunodeficiency Virus Type 1
keywords: Fed-batch cultivation; Plackett–Burman design; multiparameter optimization; scFv anti-p17; secreted recombinant protein; sequential simplex optimization