Phage display mediated immuno-PCR.

Yong-Chao Guo; Ya-Feng Zhou; Xian-En Zhang; Zhi-Ping Zhang; Yan-Mei Qiao; Li-Jun Bi; Ji-Kai Wen; Mi-Fang Liang; Ji-Bin Zhang

doi:10.1093/nar/gkl260

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PubMed · 16682441

Phage display mediated immuno-PCR.

Yong-Chao Guo·Ya-Feng Zhou·Xian-En Zhang·Zhi-Ping Zhang·Yan-Mei Qiao·Li-Jun Bi·Ji-Kai Wen·Mi-Fang Liang·Ji-Bin Zhang

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Abstract

Immuno-PCR (IPCR) is a powerful detection technology in immunological study and clinical diagnosis due to its ultrasensitivity. Here we introduce a new strategy termed phage display mediated immuno-PCR (PD-IPCR). Instead of utilization of monoclonal antibody (mAb) and chemically bond DNA that required in the conventional IPCR, a recombinant phage particle is applied as a ready reagent for IPCR experiment. The surface displayed single chain variable fragment (scFv) and phage DNA themselves can directly serve as detection antibody and PCR template, respectively. The aim of the design is to overcome shortcoming of low detection sensitivity of scFv so as to largely facilitate the real application of scFv in immunoassay. The idea has been demonstrated by applying hantaan virus nucleocapsid protein (NP) and prion protein (PrP) as detection targets in three experimental protocols (indirect, sandwich and real-time PD-IPCR assays). The detection sensitivity was increased 1000- to 10,000-folds compared with conventional enzyme-linked immunosorbent assays (ELISAs). This proof-of-concept study may serve as a new model to develop an easy to operate, low cost and ultrasensitive immunoassay method for broad applications.

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Capsid Proteins Enzyme-Linked Immunosorbent Assay Immunoassay Immunoglobulin Variable Region Peptide Library Polymerase Chain Reaction Prions Viral Core Proteins Evaluation Study Journal Article Research Support, Non-U.S. Gov't

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Yong-Chao Guo, Ya-Feng Zhou, Xian-En Zhang, Zhi-Ping Zhang, Yan-Mei Qiao, Li-Jun Bi, Ji-Kai Wen, Mi-Fang Liang, Ji-Bin Zhang. 2006-05-08. Phage display mediated immuno-PCR.. https://doi.org/10.1093/nar%2Fgkl260

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