Development of the Double Cyclic Peptide Ligand for Antibody Purification and Protein Detection

作者：Gong, Yiyi; Zhang, Lin; Li, Jin; Feng, Shan^*; Deng, Haiteng^*

来源：Bioconjugate Chemistry, 2016, 27(7): 1569-1573.

DOI：10.1021/acs.bioconjchem.6b00170

• PHAGE DISPLAY
• EXPRESSION
• BINDING
• DESIGN
• PCNA

Development of a peptide-based affinity matrix and detection reagent is important for biomedical research and the biopharmaceutical industry. In the present work, we designed and synthesized an immunoglobin G (IgG)-binding peptide ligand, Fc-III-4C. Fc-III-4C is composed of 15 residues, and the 4 cysteine residues form 2 disulfide bonds to generate a double cyclic structure. The binding affinity of the Fc-III-4C peptide toward human IgG was determined to be 2.45 nIvI (K-d), which is higher than that of IgG with Protein A/G (Pro-A/G). Importantly, the Fc-III-4C peptide displayed high affinity to various IgGs from different species. Fc-III-4C immobilized agarose beads exhibited high stability and reusability when compared with that of the Pro-A/G-immobilized beads. The conjugate of Fc-III-4C with FITC was demonstrated to be suitable for immunofluorescence detection of proteins expressed in cells. These results demonstrate that the Fc-III-4C peptide is a useful affinity ligand for antibody purification and as a protein detection reagent.

• 出版日期2016-7
• 单位北京大学; 清华大学