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Journal of Spectroscopy
Volume 2015, Article ID 248504, 9 pages
http://dx.doi.org/10.1155/2015/248504
Research Article

A Model System for Concurrent Detection of Antigen and Antibody Based on Immunological Fluorescent Method

1Key Laboratory of Optoelectronic Chemical Materials and Devices of Ministry of Education, Jianghan University, Wuhan 430056, China
2Flexible Display Materials and Technology Co-Innovation Centre of Hubei Province, Jianghan University, Wuhan 430056, China

Received 30 January 2015; Revised 8 April 2015; Accepted 15 April 2015

Academic Editor: Rizwan Hasan Khan

Copyright © 2015 Yuan-Cheng Cao. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

This paper describes a combined antigen/antibody immunoassay implemented in a 96-well plate using fluorescent spectroscopic method. First, goat anti-human IgG was used to capture human IgG (model antigen); goat anti-human IgG (Cy3 or FITC) was used to detect the model antigen; a saturating level of model antigen was then added followed by unlabelled goat anti-human IgG (model antibody); finally, Cy3 labelled rabbit anti-goat IgG was used to detect the model antibody. Two approaches were applied to the concomitant assay to analyze the feasibility. The first approach applied FITC and Cy3 when both targets were present at the same time, resulting in 50 ng/mL of the antibody detection limit and 10 ng/mL of antigen detection limit in the quantitative measurements of target concentration, taking the consideration of FRET efficiency of 68% between donor and acceptor. The sequential approach tended to lower the signal/noise (S/N) ratio and the detection of the model antigen (lower than 1 ng/mL) had better sensitivity than the model antibody (lower than 50 ng/mL). This combined antigen/antibody method might be useful for combined detection of antigens and antibodies. It will be helpful to screen for both antigen and antibody particularly in the situations of the multiserotype and high-frequency mutant virus infections.