The usefulness of CHAPS as a non-cytotoxic stabilizing agent in purification of growth factors

Abstract

Among several detergents, a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), was found to be least cytotoxic for cultured mammalian cells. CHAPS improved the activity recovery and elution profile of crude and purified fibroblast growth factors (FGFs) during chromatographies. Diluted preparations of FGFs were stabilized by CHAPS against the loss during storage. Amino acid sequence analysis was not disturbed by CHAPS. CHAPS was removable by reversed-phase high-performance liquid chromatography. These results indicate that CHAPS is useful as a non-cytotoxic stabilizing agent in purification of various kinds of bioactive polypeptides.

Monoclonal antibodies against heparin-binding growth factor II/basic fibroblast growth factor that block its biological activity: invalidity of the antibodies for tumor angiogenesis.

“…aFGF was further purified by cation-exchange HPLC on a Protein Pak G-SP column (8.2 x 75 mm) (Nihon Waters, Tokyo). The fractions eluted from the column with 0.35 M NaCI/50 mM sodium phosphate buffer (pH 6.8)/0.1% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) (20,21), which gave a single protein peak with DNA synthesis stimulating activity (see below), were collected. On SDS/19.5% polyacrylamide gel, the reduced samples of purified bFGF and aFGF gave single bands of protein with Mrs of 18,000 and 16,000, respectively.…”

“…The protein concentrations of FGFs were determined with a BCA kit with bovine serum albumin as a standard (Pierce). For RIA, mouse and human bFGFs were partially purified by heparin-5PW HPLC (Tosoh, Tokyo) from crude extracts of mouse lung and human kidney with 0.15 M ammonium sulfate (pH 4.5) as described (20). Human kidney was obtained at autopsy (within 5 hr after death).…”

Exploring Steroidal Surfactants as Potential Drug Carriers for an Anticancer Drug Curcumin: An Insight into the Effect of Surfactants’ Structure on the Photophysical Properties, Stability, and Activity of Curcumin

“…Moreover, a detailed study is still lacking regarding the applications of this unique class of biosurfactants as possible drug carriers for curcumin under different physiological conditions. CHAPS (3-[(3-cholamidopropyl)-dimethylamino]-1-propanesulfonate), a zwitterionic derivative of cholic acid, having the properties of both the bile acid and the sulfobetaine surfactant, has emerged as one of the most important amphiphiles. − Because of its biocompatibility, neutral nature over a wide pH range, and mildness toward proteins, CHAPS-based colloids could be considered safe and efficacious carriers for drugs and cosmetics. Moreover, zwitterionic-based drug carriers are known to exhibit superior biocompatibility, anti­( bio )­fouling property, negligible immunogenicity, higher cellular uptake of chemicals and drugs, etc . , Recently, the increasing attention toward the applications of zwitterionic moieties as drug carriers and therapeutics , offers immense opportunity to exploit this bioinspired surfactant in the field of drug delivery; also, a comparison with its bile salt analogues is considered necessary for the fundamental understanding and applications.…”