METHOD OF ISOLATION OF FUSED PROTEIN CONTAINING HUMAN PROINSULIN AMINO ACID SEQUENCE Russian patent published in 1996 - IPC

FIELD: biotechnology. SUBSTANCE: method involves separation of fused protein as inclusion bodies after cell disintegration from soluble impurities by microfiltration followed by diafiltration up to protein concentration 0.4-0.45 mg/ml at pH 6.0-6.3; dissolving inclusion bodies and separation of fused protein by microfiltration followed by diafiltration with 6-8 volumes of buffer with respect to microfiltrate volume up to protein concentration 0.35-0.4 mg/ml and coupled ultrafiltration; constituent proteins purification by their precipitation at pH 5.4-5.6 or by 5-fold dilution with water followed by impurities removal and desalting by microfiltration and ultrafiltration and coupled diafiltration which is carried out by 10 volumes of buffer with respect to microfiltrate volume at pH 7-8 and further lyophilization or by addition of equal volume of 96% ethanol followed by centrifugation, evaporation and supernatant lyophilization. Method can be used for preparing human recombinant insulin and other fused proteins. EFFECT: improved method of fused protein isolation. 2 dwg