Determination of the affinity constants for phage display albumin-binding peptides

PeerJ

article

Yi-Feng Shi¹ 

[1] Department of Biotechnology, School of Biological Engineering, Dalian Polytechnic University, Dalian, Liaoning Province

关键词: Phage display;    Affinity constant;    Binding peptide;    Human serum albumin;    Phage titer;    Panning criteria;    Drug carrier;   

DOI  :  10.7717/peerj.15078

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

BackgroundPhage display technology has been established as a powerful screening approach to select ligands or peptides for binding to proteins. Despite rapid growth in the field, there has been a relative dearth of quantitative criteria to measure the effectiveness of the process of phage display screening. Since human serum albumin (HSA) has been extensively studied as a drug carrier to extend the plasma half-life of protein therapeutics, the use of phage display technology is required for identifying albumin-binding peptides as the very promising strategy of albumin-binding against albumin fusion. The construction of albumin-binding drug requires the assessment of a large quantity of HSA-binding peptide (HSA binder) candidates for conjugation with therapeutic proteins. The use of the linear epitope mapping method has allowed researchers to discover many HSA-binding peptides. However, it may be inefficient to select these peptides based on sequence identity via randomly sequencing individual phage clones from enrichment pools.MethodHere, a simple assessment method to facilitate phage display selection of HSA-binding peptides was recommended. With experimentally determined phage titer, one can calculate the specificity ratios, the recovery yields and the relative dissociation constants, which are defined as quantitative criteria for panning and characterization of the binding phage fused peptides.ResultsConsequently, this approach may not only enable more rapid and low-cost phage display screening, but also efficiently reduce pseudo-positive phages selected as HSA binders for conjugation with therapeutic proteins.

【 授权许可】

CC BY