An immunoreactor-based competitive fluoroimmunoassay for monitoring staphylococcal enterotoxin B using bioconjugated quantum dots.

Vinayaka, A. C. and Thakur, M. S. (2012) An immunoreactor-based competitive fluoroimmunoassay for monitoring staphylococcal enterotoxin B using bioconjugated quantum dots. Analyst, 137. pp. 4343-4348.

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Abstract

Extensive research on avian systems has proved hens as an alternate source for polyclonal antibody generation necessary for immunosensing applications. Herein, we present the immobilization of avian antibody raised against staphylococcal enterotoxin B (SEB) and its applicability for a competitive fluoroimmunoassay technique. White leghorn hens immunized with SEB generated high affinity antibodies with a highest yield of 3.2 mg ml�1 having affinity constant of 0.976 � 1010 M l�1. A competitive fluoroimmunoassay format was developed comprising CdTe557 as a fluorescence detector for monitoring SEB, a bacterial super-antigen. CdTe557 was bioconjugated to SEB according to the carbodiimide protocol and confirmed by absorption spectral analysis. An immunoreactor column was designed by immobilizing anti-SEB antibodies and was successfully employed as an efficient biorecognition tool. An immuno-affinity reaction involving competitive binding between free SEB and CdTe557-bioconjugated SEB for immobilized antibody was relied upon to attain assay specificity and sensitivity. It was possible to quantify SEB from 1000 to 10 ng based on the integrated fluorescence of the SEB–CdTe557 bioconjugate eluted from the immunoreactor column with a limit of detection of 8.15 ng and a regression coefficient R2 ¼ 0.9925. Thus, integration of QDs with immuno-affinity reactions revealed the versatility of nanoparticles as a potential fluorescence label for bioanalytical applications.

Item Type:	Article
Uncontrolled Keywords:	foodborne pathogens, Staphylococcus aureus, quantum dots
Subjects:	600 Technology > 01 Medical sciences > 10 Immunology
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	15 Nov 2012 06:19
Last Modified:	22 Oct 2018 09:40
URI:	http://ir.cftri.res.in/id/eprint/10997

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