Effects of reaction temperature on the quantification of proteins derived from specific raw materials using ELISA

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  • 特定原材料由来タンパク質定量における ELISA 法の反応温度に対する影響
  • トクテイ ゲン ザイリョウ ユライ タンパクシツ テイリョウ ニ オケル ELISAホウ ノ ハンノウ オンド ニ タイスル エイキョウ

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Abstract

An enzyme-linked immunosorbent assay (ELISA) is widely used to test for allergens in foods. In this study, we investigated how temperature affects the performance of the “Morinaga FASPEK ELISA II Egg” from the kit under different conditions. ELISA was performed under low-temperature conditions (approximately 5°C), room temperature conditions (approximately 20–30°C), high-temperature conditions (approximately 45°C), and ultra-high-temperature conditions (approximately 65°C). The absorbance values were measured and compared for all conditions. Our results showed that compared to the absorbance at room temperature, the absorbance showed an average decrease of 44.5% and 15.7% under low-temperature and high-temperature conditions, respectively. A calibration curve could not be established under ultra-high-temperature conditions. To verify the main factor contributing to the decrease in absorbance, especially under high-temperature conditions, ELISA was carried out by independently heating the antigen in the standard egg product, the antibody on the immobilised plate, and the enzyme-labelled antibody solution from the kit to high-temperature conditions. No decrease in absorbance was observed upon heating the standard egg product alone. However, an average decrease of 6.2% and 1.7% was observed upon heating the antibody-immobilised plate alone, and heating the enzyme-labelled antibody solution alone, respectively. Therefore, while performing ELISA, it is important to control the temperature in the laboratory, especially that of the antibody-immobilised plate.

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