Stabilization of fibroblast growth factors by a non-cytotoxic zwitterionic detergent, 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (chaps) |
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Authors: | Yushi Matuo Nozomu Nishi Yasuyoshi Muguruma Yoshino Yoshitake Yoshishige Masuda Katsuzo Nishikawa Fumio Wada |
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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 |
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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. |
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Keywords: | growth factors FGF cell culture zwitterionic detergent amino acid analysis protein sequencing |
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