Issue 1, 2017

Real-time femtomolar detection of cancer biomarkers from photoconjugated antibody–phage constructs

Abstract

Here we describe novel covalent conjugates of antibody–phage for the detection of multiple cancer biomarkers using real time immuno-polymerase chain reaction (immuno-PCR). While the conventional process of immuno-PCR utilizes DNA-conjugated antibodies, chemical modification of antibodies not only reduces antibody affinity but also creates a heterogeneous population of products. However, phage naturally encapsulate genomic DNA, which can be used as a PCR template. To produce covalently conjugated antibody–phage constructs without recombinant antibody expression or chemical modification of antibodies, we incorporated a photocrosslinkable non-canonical amino acid within an antibody-binding domain displayed on one of the phage coat proteins. To correlate antigen presence to a specific DNA sequence, the phage genomes were modified with domains that recognized specific sets of primers. The crosslinked antibody–phage conjugates were then tested in a sandwich-type immunoassay using real-time PCR where low pg ml−1 concentrations of antigen could be detected and identified from a single solution containing a mixture of three different types of cancer biomarkers.

Graphical abstract: Real-time femtomolar detection of cancer biomarkers from photoconjugated antibody–phage constructs

Supplementary files

Article information

Article type
Paper
Submitted
25 Aug 2016
Accepted
02 Oct 2016
First published
04 Oct 2016

Analyst, 2017,142, 91-97

Real-time femtomolar detection of cancer biomarkers from photoconjugated antibody–phage constructs

M. Brasino and J. N. Cha, Analyst, 2017, 142, 91 DOI: 10.1039/C6AN01904H

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