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Oligoclonal Enzyme-linked Immunosorbent Assay Capable of Determining the Major Food Allergen, Ovomucoid, Irrespective of the Degree of Heat Denaturation

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Abstract

We have recently established an enzyme-linked immunosorbent assay (ELISA) for total ovomucoid determination, irrespective of the degree of its heat denaturation, by using a monoclonal antibody (mAb) 7D specific to the carbohydrate moiety of ovomucoid (Biosci Biothechnol Biochem, 68, 2490–2497, 2004). Two novel methods have been developed to improve the ELISA. First, its sensitivity was enhanced 100 times by using an oligoclonal cocktail of mAb 7D and two other mAbs with different epitopes as a second antibody. Second, it was shown that usage of denaturing reagents such as SDS and β-mercaptoethanol for extraction was acceptable for ELISA within a range of stability of a first antibody on a solid phase. Properties of the oligoclonal sandwich ELISA system thus constructed were discussed in connection with allergen labeling.

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Abbreviations

ELISA:

enzyme-linked immunosorbent assay

H-OM:

heated OM

mAb:

monoclonal antibody

N-OM:

native OM

OM:

ovomucoid

PBS:

10 mM phosphate-buffered saline pH 7.4

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Author notes

  1. Junko Hirose

    Present address: Department of Life Style Studies, School of Human Cultures, The University of Shiga Prefecture, 522-8533, Shiga, Japan

Authors and Affiliations

  1. Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, 611-0011, Uji, Japan

    Junko Hirose & Naofumi Kitabatake

  2. Department of Food and Nutrition, Kyoto Women’s University, 605-8501, Kyoto, Japan

    Yukie Murakami-Yamaguchi, Miki Ikeda & Hiroshi Narita

Authors
  1. Junko Hirose
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  2. Yukie Murakami-Yamaguchi
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  3. Miki Ikeda
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  4. Naofumi Kitabatake
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  5. Hiroshi Narita
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Corresponding author

Correspondence to Hiroshi Narita.

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Hirose, J., Murakami-Yamaguchi, Y., Ikeda, M. et al. Oligoclonal Enzyme-linked Immunosorbent Assay Capable of Determining the Major Food Allergen, Ovomucoid, Irrespective of the Degree of Heat Denaturation . Cytotechnology 47, 145–149 (2005). https://doi.org/10.1007/s10616-005-3755-6

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  • DOI: https://doi.org/10.1007/s10616-005-3755-6

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