Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein
Objective: To purify and comprehensive identify αS1-casein from bovine caseins and prepare rabbit polyclonal antibody against αS1-casein. Method: DEAE Sepharose Fast Flow anion exchange chromatography was used to separate and purify αS1-casein. The physicochemical property (isoelectric point and pro...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | zho |
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The editorial department of Science and Technology of Food Industry
2022-08-01
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| Series: | Shipin gongye ke-ji |
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| Online Access: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021110034 |
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| author | Shengfa HE Caiyun LONG Jiao WANG Meng XIONG Jiangqiang ZHAO Yan YAN Yaping DONG Xin LI Hongbing CHEN |
| author_facet | Shengfa HE Caiyun LONG Jiao WANG Meng XIONG Jiangqiang ZHAO Yan YAN Yaping DONG Xin LI Hongbing CHEN |
| author_sort | Shengfa HE |
| collection | DOAJ |
| description | Objective: To purify and comprehensive identify αS1-casein from bovine caseins and prepare rabbit polyclonal antibody against αS1-casein. Method: DEAE Sepharose Fast Flow anion exchange chromatography was used to separate and purify αS1-casein. The physicochemical property (isoelectric point and protein content), immunological technique and mass spectrometry were used to comprehensively identify αS1-casein. Then the αS1-casein was obtained by dialysis and lyophilization, polyclonal antibody was prepared by immunizing New Zealand white rabbit with the purified αS1-casein, and the specificity of polyclonal antibody was analyzed. Result: Among the four kinds of caseins, αS1-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 αS1-casein was 94.26%, and the yield was 27.19%. The titers of sera from two rabbits inoculated with purified αS1-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 αS1-casein and high specificity polyclonal antibody against αS1-casein, providing a new train of thought for αS1-casein purification and comprehensive identification, and also providing material basis for the development of immunologic method for detection of αS1-casein. |
| first_indexed | 2024-04-13T09:11:37Z |
| format | Article |
| id | doaj.art-c054c646e146458ca963b1eaa012efa1 |
| institution | Directory Open Access Journal |
| issn | 1002-0306 |
| language | zho |
| last_indexed | 2024-04-13T09:11:37Z |
| publishDate | 2022-08-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj.art-c054c646e146458ca963b1eaa012efa12022-12-22T02:52:52ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062022-08-01431510611410.13386/j.issn1002-0306.20211100342021110034-15Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-CaseinShengfa HE0Caiyun LONG1Jiao WANG2Meng XIONG3Jiangqiang ZHAO4Yan YAN5Yaping DONG6Xin LI7Hongbing CHEN8School of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaGanzhou Center for Disease Control and Prevention, Ganzhou 341000, ChinaSchool of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaSchool of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaSchool of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaSchool of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaSchool of Public Health and Health Management, Gannan Medical University, Ganzhou 341000, ChinaState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, ChinaState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, ChinaObjective: To purify and comprehensive identify αS1-casein from bovine caseins and prepare rabbit polyclonal antibody against αS1-casein. Method: DEAE Sepharose Fast Flow anion exchange chromatography was used to separate and purify αS1-casein. The physicochemical property (isoelectric point and protein content), immunological technique and mass spectrometry were used to comprehensively identify αS1-casein. Then the αS1-casein was obtained by dialysis and lyophilization, polyclonal antibody was prepared by immunizing New Zealand white rabbit with the purified αS1-casein, and the specificity of polyclonal antibody was analyzed. Result: Among the four kinds of caseins, αS1-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 αS1-casein was 94.26%, and the yield was 27.19%. The titers of sera from two rabbits inoculated with purified αS1-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 αS1-casein and high specificity polyclonal antibody against αS1-casein, providing a new train of thought for αS1-casein purification and comprehensive identification, and also providing material basis for the development of immunologic method for detection of αS1-casein.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021110034cow’s milk allergenαs1-caseinpurificationidentificationpolyclonal antibody |
| spellingShingle | Shengfa HE Caiyun LONG Jiao WANG Meng XIONG Jiangqiang ZHAO Yan YAN Yaping DONG Xin LI Hongbing CHEN Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein Shipin gongye ke-ji cow’s milk allergen αs1-casein purification identification polyclonal antibody |
| title | Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein |
| title_full | Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein |
| title_fullStr | Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein |
| title_full_unstemmed | Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein |
| title_short | Purification, Identification and Polyclonal Antibody Development for Cow’s Milk Major Allergen αS1-Casein |
| title_sort | purification identification and polyclonal antibody development for cow s milk major allergen αs1 casein |
| topic | cow’s milk allergen αs1-casein purification identification polyclonal antibody |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021110034 |
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