本文選題：花生　切入點：河蝦　出處：《江南大學》2009年碩士論文

【摘要】： 花生、河蝦屬于八大類容易引起過敏的食品,相對其它食品過敏而言,它們的過敏發(fā)生率較高、臨床癥狀更為嚴重,在公共衛(wèi)生和食品安全領域倍受人們的關注。而現(xiàn)有的過敏原檢測方法操作繁瑣,無法滿足快速、簡便、在線檢測的要求,因此,研究和建立一種方便、安全和快速的檢測方法具有重要意義。 研究主要內容包括花生、河蝦過敏原的分離純化,兔抗花生、河蝦多克隆抗體的制備,探索膠體金是否可以作為一種新型的免疫佐劑;運用自組裝的壓電免疫傳感器測定花生抗體和河蝦抗體的免疫親和常數,并同時包被兩種抗體,對過敏原分別進行篩檢。 花生、河蝦蛋白浸液經硫酸銨分級沉淀,等電點沉淀,AKTA蛋白純化系統(tǒng)得到純化,收集后經電泳檢驗。采用檸檬酸三鈉還原法制備得到16nm左右的膠體金,以新西蘭大白兔為模式動物制備抗體,免疫過程設立膠體金為佐劑和弗氏完全(不完全)佐劑作對照,發(fā)現(xiàn)膠體金能夠顯著提高大白兔體內抗花生、河蝦過敏蛋白的抗體水平,花生與膠體金組的平均效價比弗氏佐劑組提高了一倍,其IC_(50)為34.28 ng/mL;河蝦與膠體金組的平均效價比弗氏佐劑組提高了25%,其IC_(50)為43.95 ng/mL。 運用自組裝的壓電免疫傳感器測定得到花生抗體的親和常數Kα=2.86×10~7 L/mol,混合抗體對花生抗原的Kα=1.37×10~7 L/mol;河蝦抗體的Kα=5.16×10~7 L/mol,混合抗體對河蝦抗原的Kα=2.57×10~7 L/mol。自組裝的壓電傳感器同時包被花生和河蝦混合抗體,分別加入一系列濃度的花生過敏原及河蝦過敏原進行檢測,繪制得到各自的標準曲線和其檢測靈敏度。同時進行加標回收實驗,回收率較高,重現(xiàn)性較好,實現(xiàn)了花生、河蝦過敏原的同步篩檢。 本研究所建立的自組裝的壓電傳感器同時包被花生、河蝦兩種抗體,實現(xiàn)了同步對花生、河蝦過敏原的檢測,快速、方便,具有較好的實用價值。
[Abstract]:However, the existing allergen detection methods are cumbersome and can not meet the requirements of rapid, simple and on-line detection. Therefore, it is of great significance to study and establish a convenient, safe and rapid detection method.The main contents of this study include the isolation and purification of peanut and shrimp allergen, the preparation of rabbit polyclonal antibody against peanut and river shrimp, and the exploration of whether colloidal gold can be used as a new immune adjuvant.Self-assembled piezoelectric immunosensor was used to detect the immune affinity constants of peanut antibody and shrimp antibody.The protein extracts of peanut and shrimp were purified by ammonium sulfate precipitation and isoelectric point precipitation.16nm colloidal gold was prepared by tri-sodium citrate reduction method, antibody was prepared from New Zealand white rabbits, and colloidal gold was used as adjuvant and Freund's complete (incomplete) adjuvant was used as contrast in immune process.The self-assembled piezoelectric sensor was coated with mixed antibodies of peanut and prawn, and a series of peanut allergens and shrimp allergens were added to detect them, and their standard curves and sensitivity were drawn.The recovery rate was high and the reproducibility was good. The simultaneous screening of peanut and shrimp allergens was realized.
【學位授予單位】：江南大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R392.1

【引證文獻】

相關期刊論文 前1條

1 孫秀蘭;管露;單曉紅;張銀志;;食品過敏原體外檢測方法研究進展[J];東北農業(yè)大學學報;2012年02期

，

本文編號：1727118