High-Speed Gel Filtration of Polypeptides in Some Denaturants

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Abstract
The separation and analysis of proteins and polypeptides by use of a silica-based gel packing, G3000SW, for high-speed gel filtration are investigated. The peaks of bovine serum albumin, pepsin, trypsinogen, myoglobin and cytochrome c were completely separated in the presence of 0.2% SDS and 0.2 M sodium phosphate buffer (pH 7.0). The elution positions of native proteins, polypeptides in 8 M urea and polypeptide-SDS complexes were influenced by the concentration of sodium phosphate in eluents, though those of polypeptides in 6 M guanidine hydrochloride were little. These facts suggest the presence of the electrostatic interactions between negatively charged gel surfaces of the packing and polypeptides. Taking into account of the interactions, it is shown that the high-speed gel filtration by use of this column is available to the rapid estimation of molecular weight of polypeptides in both systems of SDS and 6 M guanidine hydrochloride.