中华绒螯蟹致敏蛋白的分离鉴定与纯化
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- 英文论文题名：Study on extraction,identification and purification of Chinese Mitten Crab allergen
- 论文作者：廖涛 ; 杨锴 ; 耿胜荣 ; 熊光权 ; 鉏晓艳 ; 李新 ; 江洪有 ; 程薇
- 英文论文作者：LIAO Tao ; YANG Kai ; GENG Sheng-rong ; XIONG Guang-quan ; ZU Xiao-yan ; LI Xin ; JIANG Hong-you ; CHENG Wei ; Institute of Agro-Products Processing and Nuclear-Agricultural Technology ; Hubei Academy of Agricultural Sciences ; Hubei Engineering Research Center for Farm Products Irradiation
- 年：2015
- 作者机构：湖北省农业科学院农产品加工与核农技术研究所/湖北省农产品辐照工程技术中心;
- 论文关键词：中华绒螯蟹 ; 致敏蛋白 ; 鉴定 ; 纯化
- 英文论文关键词：Chinese Mitten Crab ; allergen ; identification ; purification
- 会议召开时间：2015-09-21
- 会议录名称：中国核科学技术进展报告（第四卷）——中国核学会2015年学术年会论文集第8册（同位素分卷、辐射研究与应用分卷、核技术工业应用分卷、核农学分卷、核医学分卷、核情报分卷）
- 语种：中文
- 分类号：S917.4
- 学会代码：EGVD
- 会议名称：中国核学会2015年学术年会
- 会议地点：中国四川绵阳
- 主办单位：中国核学会
- 学会名称：中国核学会
- 页数：6
- 文件大小：385k
- 原文格式：O
- 会议级别：全国
摘要
利用十二烷基磺酸钠-聚丙烯酰胺电泳(SDS-PAGE)技术对中华绒螯蟹粗提蛋白液进行初步分离,SDS-PAGE图谱显示蟹粗提液中主要有14条深浅不一的可辨蛋白条带,分子量范围为18.6~94.5 kDa;利用中华绒螯蟹过敏患者的血清池免疫印迹分析,初步确定粗提物中能与中华绒螯蟹过敏患者血清反应的组分有9种,分子量在37.2~89.1 kDa之间。对蟹粗提蛋白经过硫酸铵分段沉淀、DEAE Sepharose CL-6B离子交换层析以及Sephadex G-100Superfine凝胶层析,逐步对中华绒螯蟹致敏蛋白进行纯化,并得到两个蛋白复合物,一个是分子量大约为75.9和89.1 kDa的蛋白复合物,另外一个是包含分子量为67.6、85.1、89.1和94.5 kDa的蛋白复合物。并利用免疫印迹实验证明了纯化后蛋白的免疫原性。
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) was applied for preliminary separation of crude protein extracts from Chinese mitten crab.The results of SDS-PAGE showed the proteins of Chinese mitten crab comprised of at least fourteen discernible protein bands with the molecular weight ranging from 18.6~94.5 kDa.The allergens were identified by Western blotting using the serum pool of patients allergic to Chinese mitten crab.There are 9 protein bands that can bind with patients' IgE,and the molecular weights ranges from 37.2~89.1 kDa.Then the crab allergen proteins were gradually purified by salting out with the ammonium sulphate,DEAE Sepharose CL-6B ion-exchange chromatography and Sephadex G-100 Superfine gel column chromatography.Two protein complexes were obtained with the molecular weight of one protein complex included 75.9 and 89.1 kDa,and the other included 67.6,85.1,89.1 and 94.5 kDa.Finally,immunogenicity of pure allergen proteins was identified by western-blotting.
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) was applied for preliminary separation of crude protein extracts from Chinese mitten crab.The results of SDS-PAGE showed the proteins of Chinese mitten crab comprised of at least fourteen discernible protein bands with the molecular weight ranging from 18.6~94.5 kDa.The allergens were identified by Western blotting using the serum pool of patients allergic to Chinese mitten crab.There are 9 protein bands that can bind with patients' IgE,and the molecular weights ranges from 37.2~89.1 kDa.Then the crab allergen proteins were gradually purified by salting out with the ammonium sulphate,DEAE Sepharose CL-6B ion-exchange chromatography and Sephadex G-100 Superfine gel column chromatography.Two protein complexes were obtained with the molecular weight of one protein complex included 75.9 and 89.1 kDa,and the other included 67.6,85.1,89.1 and 94.5 kDa.Finally,immunogenicity of pure allergen proteins was identified by western-blotting.
引文
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[6]Neil C E O,Lehrer S B.Seafood Allergy and Allergens:a review,the Institute of Food Technologists.Expert Panel on Food Safety and Nutrition,2000.
[7]Hansom L,Telemo E.The growing allergy problem.Acta Pediatr,1997,86:916-918.
[8]毛露甜,黄婉慧,王小明,等.蟹类和蟹类过敏原的交叉反应性研究[J].中国免疫学杂志,2008,24(8):748-751.
[9]Byun M W,Lee J W,Yook H S,et al.Application of gamma irradiation for inhibition of food allergy[J].Radiation Physics and Chemistry,2002,63(3):369-370.
[10]陈伟,吴兴达,刘群英.中华绒螯蟹主要变应原的纯化与应用分析[J].中国海洋药物杂志,2010,29(3):28-32.
[11]吴序栎,邓利平,刘志刚.大闸蟹过敏原的分离、鉴定与纯化[J].食品科技,2009,6:294-296.
[12]吴兴达.中华绒螯蟹主要致敏原的分离纯化与免疫学鉴定[D].2009.
[13]张华.刀额新对虾主要致敏原的分离纯化与鉴定[D].2008.
[2]吕相征,刘秀悔,杨晓光,等.健康人群食物过敏状况的初步调查[J].中国食品卫生杂志,2005,2:119-121.
[3]许岩,许红.食用蟹爬子引起过敏性休克1例[J].沈阳医学院学报,2000,2(2):108.
[4]安爱芝,孙月芹.食坑蟹引起过敏性休克1例[J].医学理论与实践,2003,16(8):87.
[5]Dean D M,Hush A S,Ronald A S.Food Alergy:Adverse reactions to foods and food additives.(Second Edition).USA Blackwell science inc 1997.
[6]Neil C E O,Lehrer S B.Seafood Allergy and Allergens:a review,the Institute of Food Technologists.Expert Panel on Food Safety and Nutrition,2000.
[7]Hansom L,Telemo E.The growing allergy problem.Acta Pediatr,1997,86:916-918.
[8]毛露甜,黄婉慧,王小明,等.蟹类和蟹类过敏原的交叉反应性研究[J].中国免疫学杂志,2008,24(8):748-751.
[9]Byun M W,Lee J W,Yook H S,et al.Application of gamma irradiation for inhibition of food allergy[J].Radiation Physics and Chemistry,2002,63(3):369-370.
[10]陈伟,吴兴达,刘群英.中华绒螯蟹主要变应原的纯化与应用分析[J].中国海洋药物杂志,2010,29(3):28-32.
[11]吴序栎,邓利平,刘志刚.大闸蟹过敏原的分离、鉴定与纯化[J].食品科技,2009,6:294-296.
[12]吴兴达.中华绒螯蟹主要致敏原的分离纯化与免疫学鉴定[D].2009.
[13]张华.刀额新对虾主要致敏原的分离纯化与鉴定[D].2008.
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