[Preparation of recombinant human insulin--study of downstream process].
This study was intended to establish a method of preparation of recombinant human insulin, with (His)6-Arg-Arg-human proinsulin (RRhPI) expressed by Escherichia coli. After DEAE-Sepharose Fast Flow ion-exchange chromatography, Sephadex G-25 chromatography and refolding, enzyme cleavage and Superdex 75 size exclusion chromatography,the RRhPI expressed by Escherichia coli in inclusion body form was converted to human insulin. The obtained recombinant human insulin was analyzed by SDS-PAGE, HPLC, amino acid composition analysis and bioidentity test (mouse convulsion test). The results indicate that our obtained preparation is highly purified, active recombinant human insulin.
Yu
Rong
R
Department of Biopharmaceutics, West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
Li
Xiaohong
X
Yang
Jiyu
J
Wu
Wutong
W
D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011384:Proinsulin / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D007328:Insulin; D011994:Recombinant Proteins; D011384:Proinsulin