Isolation of Peptide Ligands for the HIV Capsid Protein p24 by Phage-Display.

IF 2 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

International Journal of Peptide Research and Therapeutics Pub Date : 2025-01-01 Epub Date: 2025-02-07 DOI:10.1007/s10989-025-10696-0

Jerry Woo, Emily Orozco, Srinivas S Thota, Maede Chabi, Katerina Kourentzi, Richard Willson, Brian K Kay

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引用次数: 0

Abstract

Purpose: Isolate renewable and cost-efficient affinity reagents that will facilitate the detection of p24, the capsid protein of Human Immunodeficiency Virus (HIV), by screening phage-displayed combinatorial peptide libraries and identifying peptide ligands.

Method: Four in-house combinatorial peptide libraries were screened for binders in three progressive rounds against monomeric p24 protein. Peptide binders were characterized by ELISA and Surface Plasmon Resonance (SPR) and one peptide sequence was evaluated in a lateral flow assay (LFA).

Result: We identified 26 unique peptide sequences that exhibit varying phage ELISA signals above background for p24. We subsequently validated the binding of one linear and two cyclized peptide sequences with synthetic peptides. Alanine-scanning identified several residues critical to binding in the linear peptide. The linear peptide could be used for p24 detection in ELISA and LFAs.

Conclusion: Phage-displayed combinatorial peptide libraries are suitable for isolation of binders against p24 and potentially other targets. Upon identification of a minimal binding sequence, the subsequent characterization and future optimization of it can lead to a variety of diagnostic assays.

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噬菌体展示技术分离HIV衣壳蛋白p24肽配体。

目的：通过筛选噬菌体展示组合肽库和鉴定肽配体，分离出可再生且经济高效的亲和力试剂，用于检测人类免疫缺陷病毒（HIV）的衣壳蛋白p24。方法：筛选4个内部组合肽库，分三轮筛选针对p24蛋白单体的结合物。用酶联免疫吸附试验（ELISA）和表面等离子体共振（SPR）对肽结合物进行了表征，并用横向流动试验（LFA）对一个肽序列进行了评价。结果：我们鉴定出26个独特的肽序列，在p24背景上表现出不同的噬菌体ELISA信号。我们随后验证了一个线性和两个环化肽序列与合成肽的结合。丙氨酸扫描确定了几个对线性肽结合至关重要的残基。该线状肽可用于ELISA和LFAs中p24的检测。结论：噬菌体展示组合肽文库适用于分离针对p24和潜在其他靶标的结合物。在确定最小结合序列后，随后的表征和未来的优化可以导致各种诊断分析。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Peptide Research and Therapeutics 生物-生化与分子生物学

CiteScore

5.50

自引率

8.00%

发文量

131

审稿时长

>12 weeks

期刊介绍： The International Journal for Peptide Research & Therapeutics is an international, peer-reviewed journal focusing on issues, research, and integration of knowledge on the latest developments in peptide therapeutics. The Journal brings together in a single source the most exciting work in peptide research, including isolation, structural characterization, synthesis and biological activity of peptides, and thereby aids in the development of unifying concepts from diverse perspectives. The Journal invites substantial contributions in the following thematic areas: -New advances in peptide drug delivery systems. -Application of peptide therapeutics to specific diseases. -New advances in synthetic methods. -The development of new procedures for construction of peptide libraries and methodology for screening of such mixtures. -The use of peptides in the study of enzyme specificity and mechanism, receptor binding and antibody/antigen interactions -Applications of such techniques as chromatography, electrophoresis, NMR and X-ray crystallography, mass spectrometry.