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| 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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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.
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Received: 16 January 2012
Published: 25 November 2012
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[1] 易欣,柳晓磊,胡美英. 牵出技术在蛋白质相互作用研究中的应用. 生命的化学, 2011,31(2):292-297. Yi X,Liu X L,Hu M Y. Applications of the pull-down technique in the protein-protein interactions. Chemistry of Life, 2011,31(2):292-297.
[2] Vandenbulcke L,Bachett C,Van Cauwenberge P,et al. The innate immune system and its role in allergic disorders[J]. Int Arch Allergy Immunol,2006,139 (2):1592165.
[3] Jarolim E, Rumpold H, Endler A T,et al. IgE and IgG antibodies of patients with allergy to birch pollen as tools to define the allergen profile of Betula verrucosa. Allergy 1989,44:385-95.
[4] Kemeny D M, Urbanek R, Ewan P, et al. The subclass of IgG antibody in allergic disease. II.The IgG subclass of antibodies produced following natural exposure to dust mite and grass pollen in atopic and non-atopic individuals.Clin Exp Allergy, 1989,19:545-549.
[5] Svenson M, Jacobi H H, Bodtger U,et al. Vaccination for birch pollen allergy: induction of affinity-matured or blocking IgG antibodies does not account for the reduced binding of IgE to Bet v 1. Mol Immunol, 2003,39:603-612.
[6] Aalberse R C. Specific IgE and IgG responses in atopic versus nonatopic subjects. Am J Respir Crit Care Med 2000;162:124-127.
[7] Aalberse R C,Van Milligen F,Tan K Y,et al. Allergen-specific IgG4 in atopic disease. Allergy, 1993,48:559-569.
[8] Shanti K N,Martin B M,Nagpal S,et al. Identification of tropomyosin as the major shrim pallergen and characterization of its IgE binding epitopes. J Immunol, 1993, 151(10): 5354-5363.
[9] Daul C B,Slattery M,Reese G,et al. Identification of the major brown shrimp(Penaeus aztecus) allergen as the muscle protein tropomyosin. Int Arch Allergy Immunol, 1994, 105(1): 49-55.
[10] Leung P S,Chu K H,Chow W K,et al. Cloning,expression,and primary structure of metapenaeus ensis tropomyosin,the major heat-stable shrimp allergen. J Allergy Clin Immunol, 1994, 94(5): 882-890.
[11] Cao M J, Osatomi K, Hara K, et al. Identification of a myofibril-bound serine proteinase (MBSP) in the skeletal muscle of lizard fish Saurida wanieso which specifically cleaves the arginine site. Comp Biochem Physiol B Biochem Mol Biol, 2000, 125(2): 255-264.
[12] 沈关心,龚非力. 抗体技术实验指南. 北京: 科学出版社,2002. Shen G X,Gong F L. Using Antibodies:A Laboratory Manual. Beijing: Science Press,2002.
[13] 周光炎. 免疫学原理.上海:上海科学技术出版社,2007:52. Zhou G Y. Principles Of Immunology. Shanghai: Shanghai Scientific and Technical Publishers, 2007:52.
[14] 王顺涛. 抗IgE 抗体的研究进展. 细胞与分子免疫学杂志, 2004, 20(5):640-642. Wang S T. Research progress of anti-IgE antibody. Journal of Cellular and Molecular Immunology, 2004, 20(5):640-642. |
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