利用牵出技术分析与虾主要过敏原相互作用的蛋白质

中国生物工程杂志

2012, Vol. 32

Issue (11): 81-85

技术与方法

利用牵出技术分析与虾主要过敏原相互作用的蛋白质

沈梓粤, 吕哲, 秦宗华, 李任强

暨南大学生物工程学系广东省分子免疫与抗体工程重点实验室广州 510632

Applying Pull-down Technique to Study Interactional Proteins with Shrimp Allergen

SHEN Zi-yue, LV Zhe, QIN Zong-hua, LI Ren-qiang

Jinan University Department of Biotechnology, Guangdong Province Key Laboratory of Molecular Immunology and Antibody Engineering, Guangzhou 510632, China

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摘要： 目的:应用牵出技术研究与虾主要过敏原相互作用的蛋白质。方法:将纯化的虾主要过敏原原肌球蛋白偶联在用环氧氯丙烷活化的载体Sepharose-4B上作为配基并制成层析柱,以过敏原为饵蛋白从虾过敏患者血清中钓出与之相互作用的蛋白质。结果:SDS-PAGE和间接ELISA检测显示,由过敏原牵出的蛋白质至少有两种,分别是抗原特异性IgE和IgG,这说明虾过敏反应除了是由IgE所介导外,IgG也在其中起到一定的作用。结论:实验所采用的牵出技术是一个快速且有效的研究蛋白质相互作用的方法,其基于免疫亲和层析原理。

关键词： 牵出技术; 虾过敏原; 蛋白质; 相互作用

Abstract: Objective: The interactional proteins with shrimp main allergens were studied by pull-down technology. Methods: Sepharose 4B polymer beads were activated by using epichlorohydrin, shrimp main allergen (tropomyosin) was coupled to them to prepare a chromatography column, which was applied to capture the interactional proteins with shrimp main allergens from patient's serum. Results: SDS-PAGE and indirect ELISA showed that at least two proteins in patient's serum were captured by shrimp allergen, they were allergen-specific IgE and IgG respectively. The result showed that the allergy is mediated not only by IgE, IgG also plays a role. Conclusion: The pull-down technique performed is a rapid and effective method to study protein-protein interactions and is based on the immune affinity chromatography principle.

Key words: Pull-down technique Shrimp allergen Protein interaction

收稿日期: 2012-01-16 出版日期: 2012-11-25

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通讯作者: 李任强,电子信箱:trqli@jnu.edu.cn E-mail: trqli@jnu.edu.cn

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引用本文:

沈梓粤, 吕哲, 秦宗华, 李任强. 利用牵出技术分析与虾主要过敏原相互作用的蛋白质[J]. 中国生物工程杂志, 2012, 32(11): 81-85.

SHEN Zi-yue, LV Zhe, QIN Zong-hua, LI Ren-qiang. Applying Pull-down Technique to Study Interactional Proteins with Shrimp Allergen. China Biotechnology, 2012, 32(11): 81-85.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I11/81

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