期刊
ANALYST
卷 142, 期 1, 页码 91-97出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c6an01904h
关键词
-
资金
- NSF Career Award [DMR-1056808]
- NIH award [1R21EB020911-01]
Here we describe novel covalent conjugates of antibody-phage for the detection of multiple cancer bio-markers 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.
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