發(fā)布時(shí)間：2018-06-15 22:03

  本文選題：蝦過(guò)敏原 + 輻照��； 參考：《中國(guó)農(nóng)業(yè)科學(xué)院》2007年碩士論文

【摘要】： 食物過(guò)敏作為食源性疾病的一種已引起了人們的普遍關(guān)注。根據(jù)流行病學(xué)調(diào)查，美國(guó)每年約有2％～2.5％的人發(fā)生食物過(guò)敏，兒童及嬰幼兒的發(fā)病率約5％～8％。澳洲每年有600例過(guò)敏性疾病發(fā)生，并且在2004年食品企業(yè)的召回產(chǎn)品中，過(guò)敏食品比例就達(dá)47％。據(jù)不完全統(tǒng)計(jì)，我國(guó)的發(fā)病率也較高。近年來(lái)，有關(guān)食源性蝦過(guò)敏的報(bào)道屢見(jiàn)不鮮。在聯(lián)合國(guó)糧農(nóng)組織提出的八大類引起過(guò)敏的食物之中，蝦與蟹等甲殼類動(dòng)物及其制品是重要的一類。最近韓國(guó)、德國(guó)以及加拿大等國(guó)報(bào)道，輻照技術(shù)可促進(jìn)過(guò)敏原蛋白質(zhì)的降解、交聯(lián)和分子構(gòu)象的改變，破壞其抗原決定簇，有可能成為降低食物過(guò)敏反應(yīng)的技術(shù)手段。為了探索過(guò)敏原輻照脫敏的機(jī)理，本研究以中國(guó)海蝦過(guò)敏原TM(Tropomyosin)為研究對(duì)象，對(duì)不同存在狀態(tài)的過(guò)敏原(提取液、凍干粉和全蝦)做輻照處理，分析其輻照的生化效應(yīng)；利用電泳技術(shù)純化的微量過(guò)敏原免疫動(dòng)物制備高效價(jià)高特異性抗體，在此基礎(chǔ)上分析其輻照免疫學(xué)效應(yīng)，探討過(guò)敏原的輻照免疫學(xué)分子機(jī)理，為我國(guó)食品輻照脫敏技術(shù)的研究提供理論依據(jù)。 蝦是引起過(guò)敏反應(yīng)的重要食物之一，研究發(fā)現(xiàn)原肌球蛋白(Tropomyosin,TM)是蝦中一種主要過(guò)敏原，這是一種分子量為36kDa的熱穩(wěn)定性蛋白。據(jù)Lee YK和Myung-Woo Byun等報(bào)道輻照可破壞其抗原表位，降低其致敏性。本研究利用過(guò)敏原的熱穩(wěn)性及低鹽離子溶液中可溶性，成功地對(duì)過(guò)敏原作了常規(guī)提純。在此基礎(chǔ)上研究了不同劑量的輻照處理對(duì)不同存在狀態(tài)的過(guò)敏原TM的分子結(jié)構(gòu)、疏水性、濁度等生化性質(zhì)的影響，證實(shí)輻照改變了過(guò)敏原TM水溶性、疏水性，過(guò)敏原分子發(fā)生了解聚、交聯(lián)等反應(yīng)，進(jìn)而改變了其生物學(xué)特性。 為了進(jìn)一步了解輻照對(duì)過(guò)敏原致敏性的影響，在比較利用不同方法提純的過(guò)敏原蛋白質(zhì)做免疫原免疫動(dòng)物制備抗體的優(yōu)劣前提下，設(shè)計(jì)利用SDS-PAGE純化的蝦過(guò)敏原(TM)成功地制備出過(guò)敏原TM的高效價(jià)高特異性多克隆抗體，結(jié)合人血清過(guò)敏原抗體IgE，研究了不同劑量的輻照處理對(duì)不同存在狀態(tài)的過(guò)敏原與患者特異性血清抗體IgE及多抗的親和性的影響。結(jié)果發(fā)現(xiàn)：沸水處理后過(guò)敏原溶液與患者特異性血清抗體IgE仍具有很強(qiáng)的結(jié)合力，這證實(shí)了前人關(guān)于TM具有很強(qiáng)的熱穩(wěn)定性報(bào)道；蝦過(guò)敏原(TM)經(jīng)輻照處理后，TM與患者特異性血清抗體IgE及鼠源多抗(IgG)的免疫親和力隨著輻照劑量的增加而降低，表明輻照改變了過(guò)敏原的生化特性及抗原性，致敏性降低，并發(fā)現(xiàn)TM的溶液狀態(tài)要比固體狀態(tài)和活體狀態(tài)對(duì)輻照處理敏感。 研究證明輻照改變了過(guò)敏原TM的空間構(gòu)象，從而改變了過(guò)敏原的水溶性、疏水性等生物學(xué)特性，進(jìn)而使過(guò)敏原分子發(fā)生解聚、交聯(lián)、裂解等化學(xué)反應(yīng)，從而改變或者破壞過(guò)敏原抗原表位，降低了TM的致敏性。
[Abstract]:Food allergy as a food-borne disease has aroused widespread concern. According to an epidemiological survey, about 2. 5% of people in the United States develop food allergies each year, and about 5% of children and infants develop food allergies. There are 600 cases of allergic diseases in Australia each year, and the proportion of allergic food products recalled by food companies reached 47% in 2004. According to incomplete statistics, the incidence of disease in China is also high. In recent years, food-derived shrimp allergies are frequently reported. Among the eight allergenic foods proposed by FAO, crustaceans such as shrimps and crabs and their products are important. Recently, Korea, Germany and Canada reported that irradiation technology can promote the degradation of allergen proteins, cross-linking and molecular conformation changes, destroy its antigenic determinants, and may become a technical means to reduce food allergic reactions. In order to explore the mechanism of irradiation desensitization of allergen, the allergen (TMN Tropomyosin) was studied in this study. The allergens (extract, freeze-dried powder and whole shrimp) were treated by irradiation, and the biochemical effects of irradiation were analyzed. High titer and high specific antibodies were prepared by immunizing animals with microallergens purified by electrophoretic technique. On the basis of this, the effects of irradiation immunology were analyzed, and the molecular mechanism of irradiation immunology of allergens was discussed. It provides a theoretical basis for the research of food irradiation desensitization technology in China. Shrimp is one of the most important foods causing allergic reactions. Tropomyosin TMTM is a major allergen in shrimp, which is a 36 kDa thermostable protein. Lee YK and Myung-Woo Byun have reported that irradiation can destroy their epitopes and reduce their sensitivities. In this study, the allergy was successfully purified by using the thermostability of the allergen and the solubility of the allergen in low salt ion solution. On this basis, the effects of different doses of irradiation on the molecular structure, hydrophobicity, turbidity and other biochemical properties of allergen TM in different states were studied. It was proved that irradiation changed the water solubility and hydrophobicity of allergen TM. The depolymerization, crosslinking and other reactions of allergen molecules have changed their biological characteristics. In order to understand the effect of irradiation on allergen sensitivities, we compared the advantages and disadvantages of using different methods to purify allergen proteins to prepare antibodies in immunogenic immunized animals. The high titer and high specificity polyclonal antibody of allergen TM was successfully prepared by SDS-PAGE purified shrimp allergen TMN. Combined with human serum allergen antibody IgE, the effects of different doses of irradiation on the affinity of allergens in different states to patients' specific serum antibody IgE and polyantibodies were studied. The results showed that: after boiling water treatment, the allergen solution still had strong binding ability with patient specific serum antibody IgE, which confirmed that previous reports on TM had strong thermal stability. The immunological affinity of TM with specific serum antibody IgE and mouse polyantibody IgG decreased with the increase of irradiation dose, indicating that irradiation changed the biochemical characteristics and antigenicity of the allergen, and the sensitivities were decreased, and the immunological affinity of TM with the specific serum antibody IgE and mouse polyclonal antibody IgG) decreased with the increase of irradiation dose, which indicated that irradiation changed the biochemical characteristics and antigenicity of the allergen. It is also found that TM solution state is more sensitive to irradiation than solid state and living state. It has been proved that irradiation changes the spatial conformation of allergen TM, thus changes the biological properties of allergen, such as water solubility and hydrophobicity, thus causing chemical reactions such as depolymerization, crosslinking and cracking of allergen molecules. Thus, the allergen epitopes were changed or destroyed, and the sensitivities of TM were decreased.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 李兵;蟹類過(guò)敏原的BALB/c小鼠模型研究[D];集美大學(xué);2010年

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本文編號(hào)：2023798