Detection of small molecule diagnostic markers with phage-based open-sandwich immuno-PCR

作者：Dong Jinhua; Hasan Sharif; Fujioka Yuuichiro; Ueda Hiroshi^*

来源：Journal of Immunological Methods, 2012, 377(1-2): 1-7.

DOI：10.1016/j.jim.2012.01.005

• Open sandwich ELISA
• Phage display
• Real-time PCR

Open sandwich immunoassay (OS-IA) utilizes antigen-dependent stabilization of antibody variable region to quantify various antigens, enabling noncompetitive detection of small molecules with a broad working range. To further improve its detection sensitivity, here we employed phage-based immuno-PCR approach. wherein OS-IA and quantitative PCR methodologies were combined with the use of immobilized V-L fusion protein and filamentous phages displaying V-H fragment, whose DNA was extracted for PCR amplification. This approach significantly enhanced the assay sensitivity for small molecule antigens osteocalcin (BGP) peptide and 17beta-estradiol.

• 出版日期2012-3-30