Issue 45, 2018

Development and evaluation of an IgY based silica matrix immunoassay platform for rapid onsite SEB detection

Abstract

The present study involves immunoassay platform development based on a surface functionalized silica matrix for rapid onsite detection of Staphylococcal enterotoxin B (SEB). Silica matrix functionalization as well as the immunoassay parameters was experimentally designed and optimized through response surface methodology (RSM). Silica surface functionalization was carried out with hydrofluoric acid (HF), ammonia, 3-aminopropyl triethoxysilane (APTES) and glutaraldehyde (GA). The RSM optimized matrix functionalization parameters for HF, ammonia, APTES and GA were determined to be 10%, 40%, 20% and 10% (V/V), respectively. Antibodies for the study were generated against recombinant SEB toxin in rabbit (anti-SEB IgG) and chicken (anti-SEB IgY). Subsequently, antibodies were immobilized on the functionalized silica matrix and were further characterized by SEM and contact angle measurements to elucidate the surface uniformity and degree of hydrophilicity. The immunoassay platform was developed with anti-SEB IgG (capturing agent) and anti-SEB IgY (revealing partner). The limit of detection (LOD) of the developed platform was determined to be 0.005 μg mL−1 and no cross-reactivity with similar toxins was observed. Upon co-evaluation with a standard ELISA kit (Chondrex, Inc) against various field isolates, the platform was found to be on par and reliable. In conclusion, the developed method may find better utility in onsite detection of SEB from resource-poor settings.

Graphical abstract: Development and evaluation of an IgY based silica matrix immunoassay platform for rapid onsite SEB detection

Supplementary files

Article information

Article type
Paper
Submitted
25 Apr 2018
Accepted
09 Jul 2018
First published
16 Jul 2018
This article is Open Access
Creative Commons BY license

RSC Adv., 2018,8, 25500-25513

Development and evaluation of an IgY based silica matrix immunoassay platform for rapid onsite SEB detection

J. Achuth, R. M. Renuka, K. Jalarama Reddy, M. S. Shivakiran, M. Venkataramana and K. Kadirvelu, RSC Adv., 2018, 8, 25500 DOI: 10.1039/C8RA03574A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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