Affinity Purification of Insulin by Peptide-Ligand Affinity Chromatography

作者：DONG Xiaoyan FU Litang YU Haiqing

来源：Transactions of Tianjin University, 2007, (05): 313-317.

• affinity chromatography
• insulin purification
• peptide ligand
• ligand design

The affinity heptapeptide(HWWWPAS)for insulin,selected from phage display library,was coupled to EAH Sepharose 4B gel and packed to a 1-mL column.The column was used for the affinity purification of insulin from protein mixture and commercial insulin preparation.It was observed that the minor impurity in the commercial insulin was removed by the affinity chromatography.Nearly 40 mg of insulin could be purified with the 1-mL affinity column.The results revealed the high specificity and capacity of the affinity column for insulin purification.Moreover,based on the analysis of the amino acids in the peptide sequence,shorter peptides were designed and synthesized for insulin chromatography.As a result,HWWPS was found to be a good alternative to HWWWPAS,while the other two peptides with three or four amino acids showed weak affinity for insulin.The results indicated that the peptide sequence of HWWWPAS was quite conservative for specific binding of insulin.

• 出版日期2007-10-15
• 单位天津大学