title: Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture.
abstract: To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins.
authors.affiliation: Embryo Technology and Stem Cell Research Center and School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
authors.forename: Sumeth
authors.initials: S
authors.lastname: Imsoonthornruksa
authors.forename: Kamthorn
authors.initials: K
authors.lastname: Pruksananonda
authors.forename: Rangsun
authors.initials: R
authors.lastname: Parnpai
authors.forename: Ruttachuk
authors.initials: R
authors.lastname: Rungsiwiwut
authors.forename: Mariena
authors.initials: M
authors.lastname: Ketudat-Cairns
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects; D051379:Mice; D041681:NIH 3T3 Cells; D016133:Polymerase Chain Reaction; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: C100666:KCB-1 protein, recombinant; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D000469:Alkaline Phosphatase