Screening and Identification of the New Human Serum Albumin-specific Binding Peptides

doi:10.13523/j.cb.20150511

China Biotechnology

2015, Vol. 35

Issue (5): 74-80 DOI: 10.13523/j.cb.20150511

Screening and Identification of the New Human Serum Albumin-specific Binding Peptides

MA Yi, ZHAO Shao-jun, HONG An

Department of Cell Biology of Jinan University, Institute of Biological Medicine of Jinan University, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Guangzhou 510632, China

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Abstract

New human serum albumin (HSA) specific binding 7-amino-acid peptides were screened and identificated by using phage display technology. The affinity of the fusion polypeptide, comprising the screened 7-amino-acid peptides and pituitary adenylate cyclase activating peptide 27 (ML-PACAP27), were quantitatively determined,so that it was determine that the screened 7 peptides (ML1 or ML2) in fusion proteins including other peptide or protein drugs still remain the high binding activity for HSA. Using M13 phage-displayed 7-mer peptide library, the affinity effect of ML1 or ML2 and HSA were preliminarily identified through enzyme-linked immunosorbent assay (ELISA) after four screening rounds and DNA sequencing of the screened clones. Affinity constants of ML1-PACAP27 or ML2-PACAP27 and HSA were assayed by surface plasmon resonance (SPR). The experimental results showed two new 7-amino-acid peptides ML1 and ML2 were obtained, and their amino acid sequence is LKSCKPL (ML1) and SLKSHAL (ML2). The ELISA results showed the screened clones containing ML1 and ML2 can bind to HSA, and the affinity effect of ML2 for HSA is higher than that of ML1. The SPR results showed the dissociation constants of ML1 and ML2 for HSA is respectively 8.1×10^-6and 3.7×10^-6mol/L,and the affinity of ML2 and HSA is higher than that of ML1. In the study, two new HSA-specifically binding 7-amino-acid peptides that were screened and identificated may be used for the fusion expression or design of HSA-conjugated molecule drugs with long-acting effect, which can provide the new tools for extending the half-life of molecule drugs and the development of the novel drugs.

Key words： Human serum albumin(HSA) Phage-display Specific binding Drug half-life

Received: 02 March 2015 Published: 25 May 2015

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Cite this article:

MA Yi, ZHAO Shao-jun, HONG An. Screening and Identification of the New Human Serum Albumin-specific Binding Peptides. China Biotechnology, 2015, 35(5): 74-80.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150511 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I5/74

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