Protocols for Screening Peptides Binding to CCN Family Proteins and Their Extended Utility

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The function of CCN family proteins is determined by their interactions with multiple cofactors that are present in the microenvironment. Therefore, determining these cofactors is critically important in understanding the molecular function of CCN family members. For this objective, a bacteriophage random peptide display library is a suitable tool. In this library, each filamentous bacteriophage is designed to display an oligopeptide of 7–20 random amino acid residues on its surface. Bacteriophage clones that possess peptides that bind to a CCN family protein are selected through several cycles of a process called biopanning or affinity selection. By determining the nucleotide sequence of the DNA that encodes the displayed peptide, the oligopeptides that specifically bind to the CCN family member can be specified. The obtained peptide sequences can be utilized to design peptide aptamers for CCN family proteins, or as a key sequence to determine new CCN family cofactor candidates in silico. Instead of a random peptide cDNA library, an antibody cDNA library from naïve lymphocytes or from B cells immunized by a CCN family protein can be used in order to obtain a highly specific CCN family detection or functional modulation tool.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages87-101
Number of pages15
DOIs
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2582
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Antibody
  • Aptamer
  • CCN family
  • Cofactor
  • Phage display

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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