【摘要】： 魚類食物是人類獲取優(yōu)質(zhì)蛋白的重要途徑,但有部分人對(duì)魚類蛋白過敏,健康受到極大威脅。有研究表明,鯉魚和鱈魚等魚類中主要過敏原是小清蛋白,因而研究鯉魚小清蛋白的抗原性將有助于預(yù)防食物過敏和生產(chǎn)低致敏性產(chǎn)品。 本研究的主要內(nèi)容包括鯉魚小清蛋白的分離純化,兔抗鯉魚小清蛋白多克隆抗體的制備及青魚、草魚、鰱魚蛋白的粗提物與鯉魚小清蛋白之間免疫交叉反應(yīng)的研究。 通過實(shí)驗(yàn),建立了以磷酸鹽緩沖液粗提結(jié)合DEAE-Sepharose Fast Flow離子交換層析分離純化鯉魚小清蛋白的方法;SDS-PAGE和PAGE鑒定結(jié)果表明,用這種方法可以獲得純度＞90%的鯉魚小清蛋白樣品。 本實(shí)驗(yàn)建立了間接ELISA檢測(cè)兔抗鯉魚小清蛋白抗血清效價(jià)的檢測(cè)方法及制備兔抗鯉魚小清蛋白多克隆抗體的方法。免疫兩只日本雄性大白兔分別獲得了間接ELISA檢測(cè)效價(jià)高達(dá)1:560,000和1:720,000的抗血清。結(jié)合間接競(jìng)爭(zhēng)ELISA實(shí)驗(yàn)對(duì)鯉魚小清蛋白的抗原性進(jìn)行了分析和評(píng)價(jià),研究結(jié)果表明,鯉魚小清蛋白具有很好的抗原性。 通過免疫印跡和間接競(jìng)爭(zhēng)ELISA實(shí)驗(yàn),證實(shí)了鯉魚小清蛋白和青魚、草魚、鰱魚粗提蛋白之間存在免疫交叉反應(yīng),發(fā)生交叉反應(yīng)的主要蛋白分子量約為12kDa。
[Abstract]:Fish food is an important way for human to obtain high-quality protein, but some people are allergic to fish protein, and their health is seriously threatened. Studies have shown that the major allergen in fish such as carp and cod is small white protein, so studying the antigenicity of carp small albumin will help to prevent food allergy and produce hypoallergenic products. The main contents of this study include the isolation and purification of carp albumin, the preparation of rabbit anti-carp albumin polyclonal antibody, and the study of immune cross-reaction between the crude extracts of carp, grass carp and silver carp proteins and carp albumin. A method for the separation and purification of carp small white protein with phosphate buffer solution and DEAE-Sepharose Fast Flow ion exchange chromatography was established. The results of SDS-PAGE and PAGE showed that the purity of carp small white protein was more than 90% by this method. An indirect ELISA method for the detection of rabbit anti-carp serum titer and preparation of rabbit anti-carp albumin polyclonal antibody was established. Two Japanese male rabbits were immunized with anti-ELISA sera with the titers of 1? 560 000 and 1? 720 000 respectively. The antigenicity of carp small white protein was analyzed and evaluated by indirect competitive ELISA assay. The results showed that carp small white protein had good antigenicity. By means of immunoblotting and indirect competitive ELISA test, it was confirmed that there was an immune cross-reaction between carp albumin and the crude protein extracted from Cyprinus Carpio, grass carp and silver carp. The molecular weight of the main cross-reaction protein was about 12kda.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李陽;劉楚怡;薛文通;;響應(yīng)面優(yōu)化鱖魚致敏蛋白的木瓜蛋白酶水解工藝[J];食品科學(xué);2012年19期

相關(guān)碩士學(xué)位論文 前1條

1 沈海旺;章魚過敏原的研究[D];集美大學(xué);2012年

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本文編號(hào)：2465339