Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen <i>α</i><sub>S1</sub>-Casein

HE Shengfa; LONG Caiyun; WANG Jiao; XIONG Meng; ZHAO Jiangqiang; YAN Yan; DONG Yaping; LI Xin; CHEN Hongbing

doi:10.13386/j.issn1002-0306.2021110034

Volume 43 Issue 15

Jul. 2022

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Science and Technology of Food Industry > 2022 > 43(15): 106-114. > DOI: 10.13386/j.issn1002-0306.2021110034

HE Shengfa, LONG Caiyun, WANG Jiao, et al. Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen α_S1-Casein[J]. Science and Technology of Food Industry, 2022, 43(15): 106−114. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021110034.

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Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen α_S1-Casein

1.
School of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, China
2.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China
3.
Ganzhou Center for Disease Control and Prevention, Ganzhou 341000, China
4.
School of Food Science & Technology, Nanchang University, Nanchang 330047, China
5.
Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, China

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Received Date: November 04, 2021
Available Online: June 16, 2022

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Abstract

Abstract

Objective: To purify and comprehensive identify α_S1-casein from bovine caseins and prepare rabbit polyclonal antibody against α_S1-casein. Method: DEAE Sepharose Fast Flow anion exchange chromatography was used to separate and purify α_S1-casein. The physicochemical property (isoelectric point and protein content), immunological technique and mass spectrometry were used to comprehensively identify α_S1-casein. Then the α_S1-casein was obtained by dialysis and lyophilization, polyclonal antibody was prepared by immunizing New Zealand white rabbit with the purified α_S1-casein, and the specificity of polyclonal antibody was analyzed. Result: Among the four kinds of caseins, α_S1-casein had the latest peak time, the largest peak area in anion exchange chromatography and the highest position in the electrophoretic diagram. Finally, the purity of α_S1-casein was 94.26%, and the yield was 27.19%. The titers of sera from two rabbits inoculated with purified α_S1-casein for five times reached 1280000 and 320000, respectively. In addition to the slight cross reaction with soybean protein (<0.25%), the antiserum showed no cross reaction with α-Lactalbumin, β-Lactoglobulin, egg white protein and peanut protein, indicating high specificity. Conclusion: This study prepared high purity α_S1-casein and high specificity polyclonal antibody against α_S1-casein, providing a new train of thought for α_S1-casein purification and comprehensive identification, and also providing material basis for the development of immunologic method for detection of α_S1-casein.
- cow’s milk allergen,
- α_S1-casein,
- purification,
- identification,
- polyclonal antibody

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References

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