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Production of a Monoclonal Antibody by Simultaneous Immunization of Staphylococcal Enterotoxin A and B

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Abstract

In this paper, a method of simultaneous immunizing BALB/c mice with staphylococcal enterotoxin (SE) A and B (SEA and SEB) to prepare a monoclonal antibody (3F2) for detecting both of SEA and SEB was developed. The results showed that antibody 3F2 had high titers against both SEA and SEB by enzyme-linked immunosorbent assay (ELISA). The sensitivities of 3F2 to SEA and SEB detected by ELISA were 133.2 and 82.5 ng/mL, respectively, and the detection limits for the two enterotoxins were about 1 ng/mL. The antibody 3F2 had high specificities and affinities to both SEA and SEB, and had no cross-reaction with SEC1, bovine serum albumin, and ovalbumin. SEs-free skimmed milk samples were spiked with different concentrations of SEA, SEB, or both of them, respectively. Average recoveries of SEA and SEB from the spiked samples were all nearly between 82% and 104%. The result suggested that one cell fusion with simultaneous immunization by multiple antigen to prepare monoclonal antibody against them was possible, simple, and economic. The monoclonal antibody could be used in simultaneous detecting multifarious SEs.

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Abbreviations

SEs:

Staphylococcal enterotoxins

SEA:

Staphylococcal enterotoxin A

SEB:

Staphylococcal enterotoxin B

SEC1 :

Staphylococcal enterotoxin C1

BSA:

Bovine serum albumin

OVA:

Ovalbumin

ELISA:

Enzyme-linked immunosorbent assay

icELISA:

Indirect competitive ELISA

PBST:

Phosphate-buffered saline containing 0.05% Tween 20

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Acknowledgements

This research was supported by the Program of National Science Foundation of China (No.30871754), the Shanghai Committee of Science and Technology, China (Grant No.07dz19508) and the Project of the National High Technology Research and Development Program of China (863 Program, No.2006AA10Z1A3 and No.2008AA10Z416).

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Authors and Affiliations

  1. Key Laboratory of Food Safety Evaluation of the Ministry of Agriculture, Wuhan, 430070, Hubei Province, People’s Republic of China

    Aiping Liu, Fusheng Chen & Xiaohong Wang

  2. National Key Laboratory of Agro-microbiology of China, Huazhong Agricultural University, Wuhan, 430070, Hubei Province, People’s Republic of China

    Fusheng Chen

  3. College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei Province, People’s Republic of China

    Bin Liang, Yongxia Zhang, Aiping Liu, Fusheng Chen & Xiaohong Wang

  4. Food Quality Inspection and Testing Center of Agricultural Ministry of China, Wuhan, 430064, Hubei Province, People’s Republic of China

    Youxiang Zhou

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  1. Bin Liang
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Correspondence to Xiaohong Wang.

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Bin Liang and Yongxia Zhang contributed equally to this work.

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Liang, B., Zhang, Y., Liu, A. et al. Production of a Monoclonal Antibody by Simultaneous Immunization of Staphylococcal Enterotoxin A and B. Appl Biochem Biotechnol 164, 831–840 (2011). https://doi.org/10.1007/s12010-011-9177-3

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