cDNA Display-Mediated Immuno-PCR (cD-IPCR): An Ultrasensitive Immunoassay for Biomolecular Detection

Methods Mol Biol. 2021:2261:307-321. doi: 10.1007/978-1-0716-1186-9_19.

Abstract

Immuno-PCR (IPCR) is a sensitive antigen detection by means of specific antibody-DNA conjugates. To ensure the successful conjugation of a protein (an antibody) with a reporter DNA, immuno-PCR method based on cDNA display (cD-IPCR) has been introduced. The cDNA display molecule is a 1:1 covalent complex of a polypeptide and its encoding cDNA at the single molecule level, which is directly used for antigen detection and subsequent qPCR. This method can be applied to detect various antigens in biological samples, if sequences of their single-domain antibodies (VHHs) or peptide aptamers are known.

Keywords: B domain protein A; Cell-free translation; Immuno-detection; Immuno-polymerase chain reaction; Puromycin linker; cDNA display; qPCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Peptide / genetics
  • Aptamers, Peptide / immunology*
  • DNA, Complementary / genetics
  • DNA, Complementary / immunology*
  • Immunoassay*
  • Immunoconjugates / genetics
  • Immunoconjugates / immunology*
  • Polymerase Chain Reaction*
  • Single-Domain Antibodies / genetics
  • Single-Domain Antibodies / immunology*
  • Staphylococcal Protein A / analysis*

Substances

  • Aptamers, Peptide
  • DNA, Complementary
  • Immunoconjugates
  • Single-Domain Antibodies
  • Staphylococcal Protein A