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Stabilization of fibroblast growth factors by a non-cytotoxic zwitterionic detergent, 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (chaps)
Authors:Yushi Matuo  Nozomu Nishi  Yasuyoshi Muguruma  Yoshino Yoshitake  Yoshishige Masuda  Katsuzo Nishikawa  Fumio Wada
Institution:(1) Department of Endocrinology, Kagawa Medical School, 1750, Miki-cho, Kita-gun, 761-07 Kagawa, Japan;(2) Department of Biochemistry, Kanazawa Medical University Uchinada, 920-02 Ishikawa, Japan
Abstract:Summary The potential usefulness of a zwitterionic detergent, 3-(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), in the stabilization of acidic and basic fibroblast growth factors (FGFs) was examined. Among several detergents, CHAPS was found to be not only non-cytotoxic but also most useful in handing the diluted preparations of FGFs. The advantages are as follows: 1) at lower concentrations than 0.01% CHAPS did not affect growth factor activity of calf serum (CS) and the growth rate of BLAB/c 3T3 cells. The primary culture of rat prostate epithelium and colony formation of NRK-49F cells were hardly influenced by CHAPS lower than 0.003%; 2) the loss of FGFs that usually occurs due to their adherence to the surface of storage containers was effectively prevented by inclusion of 0.1% CHAPS; 3) the recovery of FGFs after storage or dialysis was significantly enhanced by inclusion of 0.1% CHAPS; 4) CHAPS at lower concentrations than 0.1% does not interfere with amino acid analysis, except that Thr may be misled only when the ratio of protein/CHAPS is low; 5) amino acid sequence analysis was hardly disturbed by CHAPS up to 0.5%. These results indicate that CHAPS is useful as a stabilizing agent for various kinds of polypeptides capable of showing biological activity at a low concentration. Editor's statement Polypeptide growth factors and hormones, especially heparin-binding growth factors (FGF), are notoriously troublesome to handle in purified form at extremely low concentrations. This report demonstrates an interesting solution to this problem.
Keywords:growth factors  FGF  cell culture  zwitterionic detergent  amino acid analysis  protein sequencing
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