Method of preparing an active human neutrophil chemotactic factor polypeptide
AssigneesDainippon Pharmaceutical Co., Ltd., The United States of America as represented by the Department of Health and Human Services
InventorsMasaaki Yamada, Ryuji Furuta, Junichi Yamagishi, Kouji Matsushima
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Abstract
A simple and efficient method of preparing active recombinant human neutrophil chemotactic factor. The human neutrophil chemotactic factor (NCF) is solubilized from a transformant cell homogenate using urea or guanidine hydrochloride. The polypeptide is further purified from the solution using conventional protocols such as CM-Sepharose CL-6B, HW-55 column, etc.. By using this method, large amounts of active human NCF can be recovered and purified.
Claims10
A method for preparing by direct expression an active recombinant human neutrophil chemotactic factor polypeptide consisting of an amino acid sequence of the formula ##STR6## which comprises:
- cultivating Escherichia coli cells harboring an expression vector inclusive of a base sequence encoding said polypeptide;
- disrupting said cells to form a transformant-homogenate;
- collecting a precipitate from the transformant-homogenate;
- and
- treating the precipitate with urea or guanidine hydrochloride at a concentration sufficient to solubilize said polypeptide.
Dependencies are inferred from the claim text.