虾过敏原提取物量效关系的表征
|
摘要
虾是主要的经济甲壳类动物，也是经常报道的引起过敏反应的食用海产品之一。在虾过敏症的诊断和治疗过程中，对于虾过敏原提取物的制备、虾品种的选择以及虾过敏原提取物的贮存、量化等都尤为重要。本论文旨在建立虾过敏原的提取程序；对虾参考物进行选择；探讨储存时间和温度及冻融循环过程中虾过敏原提取物免疫原性变化以及解决方法；最后确立虾过敏原提取物活性的定量、定效、定性的表征方法。主要的结论如下：
1、虾过敏原提取物抽提方法的研究
采用七种不同的过敏原抽提液，对南美白对虾（Penaeus vannamei）的丙酮粉进行抽提，结果表明，加入DTT的1M KCl溶液提取的虾蛋白提取的蛋白浓度为12.35mg/mL；抽提出蛋白的条带有15条，并包括主要过敏原和次要过敏原；并且其与IgG结合的能力比纯水的高57.27%。因此，加入DTT的1M KCl溶液可以用于虾过敏原提取物的准备。
2、不同虾类过敏原提取物免疫原性的研究
选取不同的虾品种，对其抽提物的蛋白组成和免疫活性进行比较。发现南美白对虾的过敏原提取物中有15条蛋白条带，且与10位患者血清反应的免疫条带一共有43条；与其他虾品种的交叉反应率最高可达到87.78%。因此，确定南美白对虾是免疫活性最强且包含全部已报道致敏蛋白的品种，可作为过敏原蛋白的规范化参考物。
3、储存过程中虾过敏原提取物活性变化的研究
选取南美白对虾为研究对象，通过测定五周内蛋白的浓度变化、蛋白组成以及活性比较，来探讨过敏原提取物的最佳储存条件。实验结果表明，虾过敏原提取物在-20℃和-80℃储存五周时，主要过敏原条带不会发生降解；提取物的过敏活性保持不变；且蛋白浓度仅分别下降5.71%和5.00%。
4、冻融循环过程中虾过敏原提取物活性变化的研究
采用南美白对虾为研究对象，通过对比不同温度不同冻融次数下的蛋白浓度、组成以及活性变化，发现冻融循环五次时，-3℃，-20℃，-80℃的蛋白浓度分别降低24.21%、30.03%和33.08%；-80℃和-20℃的过敏原冻融循环三次时产生新的56KDa的蛋白条带，而-3℃冻融循环四次时才产生这条新的蛋白条带；且在-3℃，-20℃，-80℃冻融循环五次时，其活性分别下降52.37%、66.09%和21.31%。因此，提出过敏原检测试剂在应用时，应尽量减少冻融次数以确保实验结果的准确性。
5、抗冻剂的使用对虾过敏原提取物耐储性的影响
抗冻剂的使用对蛋白的浓度、组成和活性的保持均具有明显作用，且蔗糖和海藻糖的效果好于甘油，蔗糖和海藻糖的浓度在10%和20%时都能维持较好的效果。且在-20℃、-40℃和-80℃储存时添加抗冻剂，要比室温和0℃时添加的效果好。
6、抗酶解能力表征虾过敏原提取物的研究
以不同虾品种的过敏原提取物和不同提取液抽提的虾过敏原抽提物为研究对象，通过对比胃蛋白酶酶解前后虾过敏原抽提物的活性以及抗酶解的时间，得出蛋白提取物的抗酶解能力与蛋白跟IgE抗体结合的能力不相关的结论，即抗酶解能力的测定表征过敏原的活性不可行。
7、RBL-2H3细胞释放β-己糖胺酶表征虾过敏原提取物活性的研究
10ng/mL的蛋白提取物的浓度是激发细胞产生β-己糖胺酶的最佳浓度。不同虾品种过敏原提取物，激发RBL-2H3细胞产生β-己糖胺酶的量不相同，蛋白提取物激发细胞释放β-己糖胺酶的量和β-己糖胺酶释放率之间存在着线性关系，并确定南美白对虾的过敏原提取物的活性为6.09。
本文通过虾过敏原提取物的定量、定效和定性研究，确立了过敏原提取物的提取及使用程序，建立了过敏原提取物的活性表征方法，为过敏原提取物的进一步应用建立了基础。
This paper based on the characterization of quantitative and qualitativerelationship of shrimp allergen extracts. Firstly, the procedure of shrimp allergenextraction is established and the standardization reference of shrimp allergen extract isselected. Secondly, the immunogenicity of shrimp allergen extracts in the storageprocess and freeze-thaw cycles are researched, the effect of antifreeze in allergenextracts is compared. At last, the method of allergen extracts immunogenicitycharacterization is investigated. The main results are as follows:
1. The selection of shrimp allergen extraction solution
The acetone powder of Penaeus vannamei is extracted with seven existing ormodified extracting solutions. The means of protein concentration, SDS-PAGE,indirect ELISA and immunoblotting are compared. It shows that the shrimp proteinextracted by1M KCl with DTT has high concentration and immunogenicity,including major and minor allergens.
2. The immunogenicity of different crustacean allergen extracts
Seven crustaceans which are extensively consumed in Qingdao are chose. Thecomponents of seven crustaceans are analyzed by SDS-PAGE and their allergenactivity is compared by indirect ELISA and Western blotting. The results show thatPenaeus vannamei allergen extracts have comparatively large immunogenicity. It canbe used as the reference of shrimp allergen extracts.
3. The immunogenicity of shrimp allergen extracts in the storage process
For the purpose of explore the best storage conditions of the allergen extracts,protein concentration is determined by BCA; protein composition is evaluated bySDS-PAGE electrophoresis; allergens immunogenicity is compared by indirectELISA. The results show that when shrimp allergen extracts are stored at-20℃and-80℃, the major allergen bands don’t degraded in less than five weeks, and theallergy immunogenicity of the extract remained unchanged.
4. The immunogenicity of shrimp allergen extracts in the freeze-thaw cyclesprocess
The freeze-thaw cycles effect on immunogenicity of Penaeus vannamei allergenextracts are studied. The results show that the protein concentration decreased by33.1%,30.0%and24.2%respectively when shrimp allergen extract in the cycle offreezing and thawing for five times at-80℃,-20℃and-3℃. Therefore, when theallergens protein are stored, the cycles of freeze-thawing should be reduced to ensurethe immunogenicity of shrimp didn’t change, then at-80℃and-20℃within fourfreeze-thaw cycles and-3℃less than two freeze-thaw cycles are allowed.
5. The effect of store endurance to allergen extracts with antifreeze
The usage of antifreeze can retain the concentration, composition andimmunogenicity of proteins. The results demonstrate that the use of sucrose andtrehalose are better than the glycerine. The sucrose and trehalose concentration of10%and20%could maintain anti-freezing effect. With the temperature of-20℃、-40℃and-80℃, the use of antifreeze is much more better than the room temperatureand0℃.
6. The anti-enzymolysis of allergen extracts immunogenicity
The stability of different crustacean allergen extracts and Penaeus vannameiallergen extracts with different extracting solutions in vitro simulated gastric fluid (SGF) were studied. With the comparison of the immunogenicity and the enzymolysistime of allergen extracts, the conclusion is that the stability of allergen extracts invitro simulated gastric fluid (SGF) used to be the characterization of allergen extractsimmunogenicity is unfeasible.
7、The characterization of shrimp allergen extraction activity by the RBL-2H3released β-hexosaminidase
The result show that10ng/mL is the best allergen extracts concentration toprovoke cells releasing β-hexosaminidase. Different crustacean allergen extracts withthe same concentration could make different β-hexosaminidase release. The dilutionreaching the half maximal release was calculated (ED_(50)). The reciprocal value of theED50was defined as the biological potency of Penaeus vannamei allergen extracts.
1、虾过敏原提取物抽提方法的研究
采用七种不同的过敏原抽提液，对南美白对虾（Penaeus vannamei）的丙酮粉进行抽提，结果表明，加入DTT的1M KCl溶液提取的虾蛋白提取的蛋白浓度为12.35mg/mL；抽提出蛋白的条带有15条，并包括主要过敏原和次要过敏原；并且其与IgG结合的能力比纯水的高57.27%。因此，加入DTT的1M KCl溶液可以用于虾过敏原提取物的准备。
2、不同虾类过敏原提取物免疫原性的研究
选取不同的虾品种，对其抽提物的蛋白组成和免疫活性进行比较。发现南美白对虾的过敏原提取物中有15条蛋白条带，且与10位患者血清反应的免疫条带一共有43条；与其他虾品种的交叉反应率最高可达到87.78%。因此，确定南美白对虾是免疫活性最强且包含全部已报道致敏蛋白的品种，可作为过敏原蛋白的规范化参考物。
3、储存过程中虾过敏原提取物活性变化的研究
选取南美白对虾为研究对象，通过测定五周内蛋白的浓度变化、蛋白组成以及活性比较，来探讨过敏原提取物的最佳储存条件。实验结果表明，虾过敏原提取物在-20℃和-80℃储存五周时，主要过敏原条带不会发生降解；提取物的过敏活性保持不变；且蛋白浓度仅分别下降5.71%和5.00%。
4、冻融循环过程中虾过敏原提取物活性变化的研究
采用南美白对虾为研究对象，通过对比不同温度不同冻融次数下的蛋白浓度、组成以及活性变化，发现冻融循环五次时，-3℃，-20℃，-80℃的蛋白浓度分别降低24.21%、30.03%和33.08%；-80℃和-20℃的过敏原冻融循环三次时产生新的56KDa的蛋白条带，而-3℃冻融循环四次时才产生这条新的蛋白条带；且在-3℃，-20℃，-80℃冻融循环五次时，其活性分别下降52.37%、66.09%和21.31%。因此，提出过敏原检测试剂在应用时，应尽量减少冻融次数以确保实验结果的准确性。
5、抗冻剂的使用对虾过敏原提取物耐储性的影响
抗冻剂的使用对蛋白的浓度、组成和活性的保持均具有明显作用，且蔗糖和海藻糖的效果好于甘油，蔗糖和海藻糖的浓度在10%和20%时都能维持较好的效果。且在-20℃、-40℃和-80℃储存时添加抗冻剂，要比室温和0℃时添加的效果好。
6、抗酶解能力表征虾过敏原提取物的研究
以不同虾品种的过敏原提取物和不同提取液抽提的虾过敏原抽提物为研究对象，通过对比胃蛋白酶酶解前后虾过敏原抽提物的活性以及抗酶解的时间，得出蛋白提取物的抗酶解能力与蛋白跟IgE抗体结合的能力不相关的结论，即抗酶解能力的测定表征过敏原的活性不可行。
7、RBL-2H3细胞释放β-己糖胺酶表征虾过敏原提取物活性的研究
10ng/mL的蛋白提取物的浓度是激发细胞产生β-己糖胺酶的最佳浓度。不同虾品种过敏原提取物，激发RBL-2H3细胞产生β-己糖胺酶的量不相同，蛋白提取物激发细胞释放β-己糖胺酶的量和β-己糖胺酶释放率之间存在着线性关系，并确定南美白对虾的过敏原提取物的活性为6.09。
本文通过虾过敏原提取物的定量、定效和定性研究，确立了过敏原提取物的提取及使用程序，建立了过敏原提取物的活性表征方法，为过敏原提取物的进一步应用建立了基础。
This paper based on the characterization of quantitative and qualitativerelationship of shrimp allergen extracts. Firstly, the procedure of shrimp allergenextraction is established and the standardization reference of shrimp allergen extract isselected. Secondly, the immunogenicity of shrimp allergen extracts in the storageprocess and freeze-thaw cycles are researched, the effect of antifreeze in allergenextracts is compared. At last, the method of allergen extracts immunogenicitycharacterization is investigated. The main results are as follows:
1. The selection of shrimp allergen extraction solution
The acetone powder of Penaeus vannamei is extracted with seven existing ormodified extracting solutions. The means of protein concentration, SDS-PAGE,indirect ELISA and immunoblotting are compared. It shows that the shrimp proteinextracted by1M KCl with DTT has high concentration and immunogenicity,including major and minor allergens.
2. The immunogenicity of different crustacean allergen extracts
Seven crustaceans which are extensively consumed in Qingdao are chose. Thecomponents of seven crustaceans are analyzed by SDS-PAGE and their allergenactivity is compared by indirect ELISA and Western blotting. The results show thatPenaeus vannamei allergen extracts have comparatively large immunogenicity. It canbe used as the reference of shrimp allergen extracts.
3. The immunogenicity of shrimp allergen extracts in the storage process
For the purpose of explore the best storage conditions of the allergen extracts,protein concentration is determined by BCA; protein composition is evaluated bySDS-PAGE electrophoresis; allergens immunogenicity is compared by indirectELISA. The results show that when shrimp allergen extracts are stored at-20℃and-80℃, the major allergen bands don’t degraded in less than five weeks, and theallergy immunogenicity of the extract remained unchanged.
4. The immunogenicity of shrimp allergen extracts in the freeze-thaw cyclesprocess
The freeze-thaw cycles effect on immunogenicity of Penaeus vannamei allergenextracts are studied. The results show that the protein concentration decreased by33.1%,30.0%and24.2%respectively when shrimp allergen extract in the cycle offreezing and thawing for five times at-80℃,-20℃and-3℃. Therefore, when theallergens protein are stored, the cycles of freeze-thawing should be reduced to ensurethe immunogenicity of shrimp didn’t change, then at-80℃and-20℃within fourfreeze-thaw cycles and-3℃less than two freeze-thaw cycles are allowed.
5. The effect of store endurance to allergen extracts with antifreeze
The usage of antifreeze can retain the concentration, composition andimmunogenicity of proteins. The results demonstrate that the use of sucrose andtrehalose are better than the glycerine. The sucrose and trehalose concentration of10%and20%could maintain anti-freezing effect. With the temperature of-20℃、-40℃and-80℃, the use of antifreeze is much more better than the room temperatureand0℃.
6. The anti-enzymolysis of allergen extracts immunogenicity
The stability of different crustacean allergen extracts and Penaeus vannameiallergen extracts with different extracting solutions in vitro simulated gastric fluid (SGF) were studied. With the comparison of the immunogenicity and the enzymolysistime of allergen extracts, the conclusion is that the stability of allergen extracts invitro simulated gastric fluid (SGF) used to be the characterization of allergen extractsimmunogenicity is unfeasible.
7、The characterization of shrimp allergen extraction activity by the RBL-2H3released β-hexosaminidase
The result show that10ng/mL is the best allergen extracts concentration toprovoke cells releasing β-hexosaminidase. Different crustacean allergen extracts withthe same concentration could make different β-hexosaminidase release. The dilutionreaching the half maximal release was calculated (ED_(50)). The reciprocal value of theED50was defined as the biological potency of Penaeus vannamei allergen extracts.
引文
[1] Standardization of Allergen Extracts，Jrgen Nedergaard Larsen and Sten Dreborg. Methodsin Molecular Medicine: Allergy Methods and Protocols, M. G. Jones and P. LympanyHumana Press Inc., Totowa, NJ,133.
[2] B.P. Singh and S.V. Gangal. Defined Allergen Extracts; Need for Efficient Diagnosis ofAllergy and Immunotherapy. Indian J Allergy Asthma Immunol.2001,15(2):67-74.
[3] Anonymous.Allergen immunotherapy:therapeutic vaccines for allergic diseases WHOposition paper. Allergy.1998,53:5-42.
[4] Anonymous. The use of standardized allergen extracts. Position statement from thecommittee on allergen standardization. American Academy of Allergy, Asthma, andImmunology(AAAAI). Allergy Clin. Immunol.1997,99:583-586.
[5] Nelson H.S. The use of standardized extracts in allergen immunotherapy. Allergy Clin.Immunol.2000,106:41-45.
[6] FAO. Report of the FAO technical consultation on food allergies. Rome1995.
[7] Daul. C. B., J. E. Morgan. and S. B. Lehrer. Hypersensitivity reactions to crustacea andmollusks. Clin. Rev. Allergy.1993,11(2):201-222.
[8] Aas K: Studies of hypersensitivity to fish. A clinical study. Int Arch Allergy Appl Immunol.1966,29:346-363.
[9] Shibaski M, Suzuki S, Tajima S, et al: Allergenicity of major component proteins ofsoybeans. Int Arch Allergy Appl Immunol.1980,61:441-448.
[10] Yunginger JW, Jones R: A review of peanut chemistry—implications for the standardizationof peanut extracts. Presented at4th Int. Paul Ehrlich Symposium on Regulatory Control andStandardization of Allergenic Extracts, Bethesda, MD,1985, October16-18.
[11] Robert F. Lemanske, Jr, and Steve L. Taylor.Standardized Extracts, Foods. CLIN REVALLERGY.1987,5:23-36.
[12] Dannaeus A, Inganas M: A follow-up study of children with food allergy. Clinical course inrelation to serum IgE-and IgG-antibody levels to milk, fish, and eggs. Clin Allergy.1981,11:533-539.
[13] Bock SA: The natural history of food sensitivity. J Allergy Clin Immunol.1982,69:173-177.
[14] Ford RPK, Taylor B: Natural history of egg hypersensitivity. Arch Dis Child.1982,57:649-652.
[15] Cherry JP, Neucere NI, Ory RL: Comparison of proteins of peanuts grown in different areas.I. Disc electrophoretic analysis of qualitative and quantitative variation. J Am Peanut ResEduc Assoc Inc.1971,3:63-74.
[16] Basha SMM: Identification of cultivar differences in seed polypeptide composition ofpeanuts (Arachis hypogaea L.) by two dimensional polyacrylamide gel electrophoresis. PlantPhysiol.1979,63:301-306.
[17] Barnett D, Baldo BH, Howden MEH: Multiplicity of allergens in peanuts. J Allergy ClinImmunol.1983,72:61-68.
[18] Arachis hypogaea L.) seed during oil roasting. J Agric Food Chem.1985,33:350-353.
[19] Perlman F: Food allergens, in Catsimpoolas N (ed): Immunological Aspects of Foods.Westport, CT, AVI Publishing Company,1977,279-316.
[20]刘锐,李冉,陈洁.我国水产品消费特征及增长潜力.农业消费展望.2011,3:53-58.
[21] FAO. Report of the FAO Technical Consultation on Food Allergies. Rome.1995.
[22] Sicherer, S.H., Munoz-Furlong, A., Sampson, H.A. Prevalence of seafood allergy in theunited states determined by a random telephone survey. J. Allergy Clin. Immunol.2004,114(1):159-165.
[23] Daul, C.B., Slattery, M., Reese, G., Lehrer, S.B. Identification of the major brown shrimp(Penaeus aztecus) allergen as the muscle protein tropomyosin. Int. Arch. Allergy Immunol.1994,105(1):49-55.
[24] Wild, L.G., Lehrer, S.B. Fish and shellfish allergy. Curr. Allergy Asthma Rep.2005,5,74-79.
[25] SAMPSON H A. Food Allergy. Part1. Immunopathogenesis and clinical disorders. J AllergyClin Immunol,1999,103(5):717-728.
[26] Daul C B,Slattery M,Reese G,et al.Identification of the major brown shrimp(Penaeusaztecus)allergen as the muscle protein tropomyosin.Int Arch Allergy ApplImmunol,1994,105:49.
[27] Hughes D A,Mills C.Food allergy:a problem on the increase. Biologist(London),2001,48(5):201-204.
[28] Musmand JJ, Daul CB, Lehrer SB. Crustacea allergy. Clin Exp Allergy.1993,23:722-32.
[29] Wang J, Sampson HA. Food anaphylaxis. Clin Exp Allergy.2007,37:65160.
[30] SAMPSON H A. Update on food allergy. Journal of Allergy and Clinical Immunology.2004,113:805-819.
[31] SAMPSON H A. Food allergy Part1: Immunopathogenesis and clinical disoders. Journal ofAllergy and Clinical Immunology.1999,103:717-728.
[32] EMMETT S E, ANGUS F J, FRY J S, et al. Perceived prevalence of peanut allergy in GreatBritain and its association with other atopic conditions and with peanut allergy in otherhouseholdmembers. Allergy.1999,54(4):380-385.
[33] MONERET-VAUTRIN D A. Epidemiologie de I allergie alimentaire et prevalence relativedes trophallergenes. Cahiers Nutrition Dietetique.2001,36(4):247-252.
[34] SCHARFER T, BOHLER E, RUHDORFER S, et al. Epidemiology of food allergy/foodintolerance in adults: associations with other manifestations of atopy. Allergy.2001,56(12):1172-1179.
[35]李振兴,林洪,李明华等.不同虾类的过敏原及其过敏原性.水产学报.2006,30(3):281-284.
[36] Goldman AS, Sellars WA, Holpern SR, et al: Milk allergy. II. Skin testing of allergic andnormal children with purified milk proteins. Pediatrics.1963,32:572-579.
[37] Lebenthal E: Cows milk protein allergy. Pediatr Clin.1975,22:827-833.
[38] Visakorpi JK, Immonen P: Intolerance to cows' milk and wheat gluten in the primarymalabsorption syndrome in infancy. Acta Pediatr Scand.1967,56:49-56
[39] Kletter B, Gery I, Freier S, et al: Immunoglobin E antibodies to milk proteins. Clin Allergy1.1971,249-255.
[40] Bleumink E, Young E: Identification of the atopic allergen in cows' milk. Int Arch AllergyAppl Immunol.1971,34:521-543.
[41] Spies JR, Stevan MA, Stein WJ, et al: The chemistry of allergens XX. New antigensgenerated by pepsin hydrolysis of bovine milk proteins, J Allergy.1970,45:208-219.
[42] Haddad ZH, Kalra V, Verma S: IgE antibodies to peptic and peptic-tryptic digests of betalactoglobulin: Significance in food hypersensitivity. Ann Allergy.1979,42:368-371.
[43] Galland A V, DoryD, Pons L. Purification of a41kD acod-allergenic protein. J Chromato B,1998,706:63-67.
[44] Van D T, Hordvik I, Endresen C. Characterization of parvalbumin, the major allergen inAlaska pollack and comparison with codfish. Mol Immuno,l.2005,42(3):345-353.
[45] Van D T, Hordvik I, Endresen C. The major allergen (parvalbumin) of codfish is encoded byat least two isotypic genes: cDNA cloning, expression and antibody binding of therecombinant allergens. Mol Immuno.2003,39(10):595-602.
[46]李振兴，林洪，李明华等.不同虾类的过敏原及其过敏原性.水产学报.2006，30(2):281-284.
[47] Chu,K.H.,Wong,S.H.,and Leung,P.S.Tropomyosin is the major mollusk allergen:reversetranscriptase polymerase chain reaction, expression and IgE reactivity. Mar Biotechnol.2000,2(5):499~509.
[48]毛露甜,范红英,张在军,黄婉慧,杨华胜,周立斌.虾类和蟹类的过敏原组分分析.细胞与分子兔疫学杂志.2008,24(6):642-644.
[49]毛露甜,黄婉慧,吴小明,范红英,张在军.虾类和蟹类过敏原的交叉反应性研究.中国免疫学杂志.2008,24:748-752.
[50] Platts-Mills T.A.E., Chapman M.D. Allergen standardization J. Allergy Clin. Immunol.1991,87:621-625.
[51] Frankland A.W. The characterization of allergens. Clin. Exp. Allergy.1996,26:865-867.
[52] Anonymous. Allergen nomenclature. WHO/IUIS Allergen nomenclature subcommittee WorldHealth Organization, Geneva, Switzerland. Clin. Exp. Allergy.1995,25:21-37.
[53] Robret E.Reisman, M.D., Review of Procedures Used in the Practice of Allergy,1981,57(7):549-558.
[54]邓鹏，孙守义，王守经，张奇志，食品过敏原的安全管理，中国食物与营养，2008,12-14.
[1] Jona,R., Fronda,A., Comparative histochemical analysis of cell wall polysaccharides byenzymatic and chemical extractions of two fruits, Biotech. Histochem.1997,72:22-28.
[2] Smith, P.K., Krohn R.I., Hermanson, G.TT.,Mallia, A.K., Gartner, F. H., Provenzano,M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. Measurement of protein usingbicinchoninic acid. Anal. Biochem.1985,150(1):76-85.
[3]张轶群,李振兴,林洪,等.果糖和木糖在美拉德反应中对虾类过敏原活性影响的研究.食品科学,2009,30(29):11-14.
[4] Motoyama K., Suma Y., Ishizaki S., Nagashima Y., Shiomi K. Molecular cloning oftropomyosins identified as allergens in six species of Crustaceans. Journal of Agriculturaland Food Chemistry,2007,55(3):985-991.
[5] Steinhoff, M.; Fischer, M.; Paschke-Kratzin, A. Comparison of extraction conditions for milkand hen's egg allergens. Food Additives&Contaminants: Part A.2011,28,(4):373-383.
[6]刘一璇,林洪,吴丽莎,等.加工方式对刀额新对虾主要过敏原免疫原性的影响.水产学报.2011,35(6):948-953.
[7] Ruegg. UT, Rudinger. J.“Reductive cleavage of cystine disulfides withtributylphosphine”. Methods Enzymol.1977,47:111-116.
[8] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V., Identification oftropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes. J.Immunol.1993,151(10):5354-5363.
[9] Motoyama, K., Suma, Y., Ishizaki, S., Nagashima, Y., Shiomi, K. Molecular cloning oftropomyosins identified as allergens in six species of crustaceans. J. Agric. Food Chem.2007,55(3):985-991.
[10] D. Wessel, U.I. Flügge. A method for the quantitative recovery of protein in dilute solution inthe presence of detergents and lipids. Analytical Biochemistry.1984,138(1):141-143.
[1] FAO/WHO. Evaluation of allergenicity of genetically modified foods, in Report of a JointFAO/WHO Expert Consultation of Allergenicity of Foods Derived from Biotechnology.World Health Organization, Geneva.2001,22-25.
[2] Lopata, A.L., Lehrer, S.B. New insights into seafood allergy. Curr Opin Allergy ClinImmunol.2009,9:270-277.
[3] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V. Identification oftropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes. J.Immunol.1993,151(10):5354-5363.
[4] Daul, C.B., Slattery, M., Reese, G., Lehrer, S.B. Identification of the major brown shrimp(Penaeus aztecus) allergen as the muscle protein tropomyosin. Int. Arch. Allergy Immunol.1994,105(1):49-55.
[5] Leung, P.S., Chu, K.H., Chow, W.K., Ansari, A., Bandea, C.I., Kwan, H.S., Nagy, S.M.,Gershwin, M.E. Cloning, expression, and primary structure of Metapenaeus ensistropomyosin, the major heat-stable shrimp allergen. J. Allergy Clin. Immunol.1994,94(5):882-890.
[6] Leung, P.S.C., Chen, Y.S., Mykles, D.L., Chow, W.K., Li, C.P., Chu, K.H. Molecularidentification of the lobster muscle protein tropomyosin, as a seafood allergen. Mol. Mar.Biol. Biotechnol.1998,7:12-20.
[7] Bauermeister, K., Wangorsch, A., Perono Garoffo, L., Reuter, A., Conti, A., Falk, S., Taylor,S.L., Vieths, S., Holzhauser, T., Ballmer-Weber, B.K., Reese, G. Novel Crustacean AllergensIdentified in North Sea Shrimp Crangon and other Crustacean Species-SarcoplasmicCalcium-binding Protein, Troponin C, Troponin I, Triosephosphate Isomerase, and MyosinLight Chain.2008, http://www.ncbi.nlm.nih.gov/protein/238477329.
[8] Shiomi, K., Sato, Y., Hamamoto, S., Mita, H., Shimakura, K. Sarcoplasmic calciumbindingprotein: identification as a new allergen of the black tiger shrimp Penaeus monodon. Int. Arch.Allergy Immunol.2008,146:91-98.
[9] Ayuso, R., Grishina, G., Ibáňez, M.D., Blanco, C., Carrillo, T., Bencharitiwong, R., Sánchez,S., Nowak-Wegrzyn, A., Sampson, H.A. Sarcoplasmic calcium-binding protein is anEF-hand-type protein identified as a new shrimp allergen. J.Allergy Clin. Immunol.2009,124:114-120.
[10] Rosal′a Ayuso, MD, PhD, Galina Grishina, MS, Ludmilla Bardina, MS, Teresa Carrillo, MD,PhD, Carlos Blanco, MD, PhD, María Dolores Ibáňez, MD, PhD, Hugh A. Sampson, MDand Kirsten Beyer, MD. Myosin light chain is a novel shrimp allergen, Lit v3. AmericanAcademy of Allergy, Asthma&Immunology.2008,122:795-802.
[11] Ayuso, R., Grishina, G., Bardina, L., Carrillo, T., Blanco, C., Ibaňez, M.D., et al Myosin lightchain is a novel shrimp allergen, lit v3. J. Allergy Clin. Immunol.2008,122(4)：795-802.
[12] Yu, C.J., Lin, Y.F., Chiang, B.L., Chow, L.P. Proteomics and immunological analysis ofanovel shrimp allergen, Pen m2. J. Immunol.2003,170:445-453.
[13] Gar a-Orozco, K.D., Aispuro-Hern′andez, E., Yepiz-Plascencia G, Calde on-de-la-Barca,A.M., Sotelo-Mundo, R.R. Molecular characterization of arginine kinase, an allergen fromthe shrimp Litopenaeus vannamei. Int. Arch. Allergy Immunol.2007,144:23-28.
[14] Smith, P.K., Krohn R.I., Hermanson, G.TT.,Mallia, A.K., Gartner, F. H., Provenzano,M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. Measurement of protein usingbicinchoninic acid. Anal. Biochem.1985,150(1):76-85.
[15]张轶群,李振兴,林洪等.果糖和木糖在美拉德反应中对虾类过敏原活性影响的研究.食品科学.2009,30(29):11-14.
[16] Leung, P.S.C., Chen, Y.S., Gershwin, M.E., Wong, S.H., Kwan, H.S., Chu, K.H.Identification and molecular characterization of Charybdis feriatus tropomyosin, the majorcrab allergen. J. Allergy Clin. Immunol.1998,102:847-852.
[17] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V. Identification oftropomyosin as the major shrimp allergen and characterization of its IgE binding epitopes. J.Immunol.1993,151(10):5354-5363.
[18] Motoyama, K., Suma, Y., Ishizaki, S., Nagashima, Y., Shiomi, K. Molecular cloning oftropomyosins identified as allergens in six species of crustaceans. J. Agric. Food Chem.2007,55(3):985-991.
[19] Bredehorst, R., David, K. What establishes a protein as an allergen? J. Chromatogr. B.Biomed Sci Appl.2001,756,33-40.
[20] Leung, P. S. C., Chow, W. K., Duffey, S., Kwan, H. S., Gershwin, M. E., Chu, K. H. IgE
reactivity against a cross-reactive allergen in crustacea and mollusca: evidence for
tropomyosin as the common allergen. J. Allergy Clin. Immunol.1996,98:954-961.
[1] Surapon Piboonpocanun, Siribangon Boonchoo, Wipada Pariyaprasert, NualanongVisitsunthorn and Orathai Jirapongsananuruk. Determination of storage conditions for shrimpextracts: analysis of specific IgE-allergen profiles. Asian Pacific Journal of Allergy andImmunology.2010,28:47-52.
[2] Jirapongsananuruk O, Sripramong C, Pacharn P, et al. Specific allergy to Penaeus monodon(seawater shrimp) or Macrobrachium rosenbergii (freshwater shrimp) in shrimp-allergicchildren. Clin Exp Allergy.2008,38:103847.
[3] Carnes J, Ferrer A, Huertas AJ, Andreu C, Larramendi CH, Fernandez-Caldas E. The use ofraw or boiled crustacean extracts for the diagnosis of seafood allergic individuals. Ann
Allergy Asthma Immunol.2007,98:34954.
[1]赵俊芳,王学谦.食物过敏原检测及其应用前景[J].中华检验医学杂志.2007,30(8):948-950.
[2]贺娟,张玉,肖会.阿罗格脱敏治疗支气管哮喘的疗效观察及护理[J].现代中西医结合杂志.2009,18(19):2336-2337.
[3]匡嘉丽,方向佳,吴军等.阿罗格长效变应原脱敏治疗变应性鼻炎的疗效观察[J].海南医学院学报.2009,15(8):883-886.
[4]张春林.阿罗格标准化变应原特异性免疫治疗对儿童过敏性哮喘的持续影响[J].药物与临床.2009,47(12):89-90.
[5] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685.
[6] MOTOYAMA K., SUMA Y., ISHIZAKI S. et al. Molecular cloning of tropomyosinsidentified as allergens in six species of Crustaceans. Journal of Agricultural and FoodChemistry.2007,55(3):985-991.
[7] WATSON P.F. The causes of reduced fertility with cryopreserved semen. AnimalReproduction Science.2000,60-61:481-492.
[8]杨继东.冻融对血浆总蛋白含量的影响[J].现代医药卫生.2004,20(3):1271.
[9] XIUFANG XIA, BAOHUA KONG, QIAN LIU, et al. Physicochemical change and proteinoxidation in porcine longissimus dorsi as inuenced by different freeze-thaw cycles. MeatScience.2009,83:239-245.
[10] SADETTIN TURHAN, N. SULE USTUN, INCI BANK. Effect of freeze-thaw cycles ontotal and heme iron contents of bonit (Sardasarda) and bluesh (Pomatomus saltator) fillets.Journal of Food Composition and Analysis.2006,19:384-387.
[11] N.S.PRATHALINGAM, S.G.REVELL, S.MIRCZUK, et al. Impact of antifreeze proteins andantifreeze glycoproteins on bovine sperm during freeze-thaw. Theriogenology.2006,66:1894-1900.
[12] STROM C S, LIU XY, JIA Z. Antifreeze protein-induced morphological modificationmechanisms linked to ice binding surface. Biological Chemistry.2004,279(31):32407-32417.
[13] SHANN-TZONG JIANG, DER-CHYAN HWANG, CHING-SAN CHEN. Effect of storagetemperature on the formation of disulfides and denaturation of milkfish actomyosin. Foodscience.1988,53(5):1333-1335.
[14]曾名勇,黄海,李八方.鳙肌肉蛋白质生化特性在冻藏过程中的变化[J].水产学报,2003,27(5):480-485.
[15]曾名勇,黄海,李八方.不同冻藏温度对鲈鱼肌肉蛋白质生化特性的影响[J].青岛海洋大学学报,2003,33(33):525-530.
[1]吴丽莎，李振兴，刘一璇，陈瑜，林洪.冻融循环过程中虾过敏原的免疫原性变化.水产学报.2011,35(5):137-141
[2] Wakamatsu T, Sato Y. Determination of unfreezable water in sucrose, sodium chlorideandprotein solutions by differential scanning calorimeter. Nippon Nogeikagaku Kaishi,1979,53：415-420.
[3] Wakamatsu T，Saito Y. Further study on the effect of sucrose and sodium chloride on gelatinand unfreezable water of egg yolk during freezing. Nippon Nogeikagaku Kaishi,1980,54:951-957.
[4] Matsumotom I,Ooizumi T,Arai K.Protective effect of sugar on freeze-denaturation of carpmyofibrillar protein. Nippon Suisan Gakkaish,1985,51:833-839.
[5]曲楠，曾名湧，董士远.海藻糖对罗非鱼糜蛋白抗冻作用的研究[J].肉类研究,2010,(1):79-82.
[6]薛勇,薛长湖,李兆杰,等.海藻糖对冻藏过程中鳙肌原纤维蛋白冷冻变性的影响[J].2006,13:637-641.
[7] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685.
[8]李振兴,林洪,李明华等.不同虾类的过敏原及其过敏原性[J].水产学报.2006,30(3):281-284.
[9] Kauzmann W. Some factors in the interpretation of protein denaturation. Protein Chemistry，1959,14:1-63.
[10] Hanafusa N.Biological macromolecule and water [C]//Japanese Society of Fisheries Science.Food and Water.Koseisha Koseikaku，Tokyo.1973:9-24.
[11]严宏忠.鱼肉蛋白质的冷冻变性及防止方法[J].中国水产,1995,(3):37-38.
[12]张娅楠,赵利,刘华,袁美兰,邓丹雯.水产品的冷冻变性及鱼糜抗冻剂研究进展.[J].河南工业大学学报(自然科学版),2011,32(6):88-92.
[1] Taylor, S.L., Lehrer, S.B. Principles and characteristics of food allergens. Crit. Rev. Food Sci.Nutr.1996,36(S):91-118.
[2] Huang, Y.Y., Liu, G.M., Cai, Q.F., Weng, W.Y., Maleki, S.J., Su, W.J., Cao, M.J. Stabilityof major allergen tropomyosin and other food proteins of mud crab (Scylla serrata) by invitro gastrointestinal digestion. Food Chem. Toxicol.2010,48,1196-1201.
[3] Astwood J D，Leach J N，Fuchs R L.Stability of food allergens to digestion in vitro. NatBiotechnol.1996,14(10):1269-1273.
[4] Kenna J G，Evans R M.Digestibility of proteins in simulated gastric fluid.Toxicologist.2000,54:141.
[5] Fu T J.Digestion stability as a criterion for protein allergenicity assessment. Ann N Y AcadSci.2002,964:99-110.
[6] Thomas, K., Aalbers, M., Bannon, G.A., Bartels, M., et al. A multi-laboratory evaluation of acommon in vitro pepsin digestion assay protocol used in assessing the safety of novelproteins. Regul. Toxicol. Pharmacol.2004,39(2):87-98
[7] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685
[8] Liu, G.M., Huang, Y.Y., Cai, Q.F., Weng, W.Y., Su, W.J., Cao, M.J. Comparative study of invitro digestibility of major allergen, tropomyosin and other proteins between Grass prawn(Penaeus monodon) and Pacific white shrimp (Litopenaeus vannamei). J. Sci. Food Agric.2011,91:163-170.
[9] Fu T J，Abbott U R，Hatzos C. Digestibility of food allergens and nonallergenic proteins insimulated gastric fluid and simulated intestinal fluid—A comparative study. J Agric FoodChem.2002,50:7154-7160.
[1] Passante E,Ehrhardt C,Sheridan H,et al.RBL-2H3cells arean imprecise model for mast cellmediator release. Inflamm Res.2009,58(9):611-618.
[2] Eccleston E，Leonard BJ，Lowe JS，et al. Basophilic leukaemia in the albino rat and ademonstration of the basopoietin. Nat New Biol.1973,44:73-76.
[3] Hoffmann, A.Jamin, K. Foetiseh, et al. Determination of the allergenic activity of birehpollen and apple prick test solutions by measurement of β-hexosaminidase release fromRBL-2H3cells: Comparison with classieal methods in allergen standardization. Allergy.1999,54:446-454.
[4] Hisashi M,Sachiko S,Toshio M,et al.Bioactive constituents from Chinese naturalmedicines—Inhibitors of antigen-induced degranulation in RBL-2H3cells from the seeds ofPsoralea corylifolia. Chem Pharm Bull.2007,55:106-110.
[5] S.Kaul, D.Luttkopf, B.Kastner, L.Vogel, G.H ltz, S.Vieths and A.Hoffmann. Mediator releaseassays based on human or murine immunoglobulin E in allergen standardization. Clinicaland Experimental Allergy.2007,37,141-150.
[6] Ward MD, Selgrade MK. Animal models for protein respiratory sensitizers. Methods.2007,(41):80-90.
[7] Susanne Kaul, Stephan Scheurer, Nicole Danz, Simone Schicktanz, Stefan Vieths,Andreas Hoffmann. Monoclonal IgE antibodies against birch pollen allergens: Noveltools for biological characterization and standardization of allergens. Allergy ClinImmunol.2003,111(6):1262-1268.
[8] Sawanee Kraithep, Kwunchit Oungbho, Supinya Tewtrakul. Anti-allergic activity ofThai medicinal plants used in longevity formulation. Songklanakarin J. Sci. Technol.2008,30(5):621-625.
[9] Tomohiro Itoh, Yasunori Fujita, Mikako Ito, Akio Masuda, Kinji Ohno, MasatoshiIchihara, Toshio Kojima, Yoshinori Nozawa, Masafumi Ito. Molecular hydrogensuppresses FcεR I-mediated signal transduction and prevents degranulation of mastcells. Biochemical and Biophysical Research Communications.2009,389(4):651-656.
[10] Eun-Kyung PARK, Min-Kyung CHOO, Eun-Jin KIM, Myung Joo HAN, Dong-HyunKIM. Antiallergic activity of ginsenoside Rh2. Biological and PharmaceuticalBulletin.2003,26(11):1581-1584.
[11] LU Y M，LI L，LUO W T，et al. A study on the change of calcium signal in mast cellactivated by antigen.Chin J Tubere Respir Dis.2000,23(4):243-246.
[12] Passante E，Frankish N. The RBL-2H3cell line: its provenance and suitability as amodel for the mast cell. Inflamm Res.2009,58(11):737-745.
[13] R.J. Dearman，R.A. Skinner，N. Deakin，et al. Evaluation of an in vitro method for themeasurement of specific IgE antibody responses: the rat basophilic leukemia (RBL) cellassay. Toxicology.2005,206:195-205.
[14] Passante E，Ehrhardt C，Sheridan H，et al. RBL-2H3cells are an imprecise model for mastcell mediator release. Inflamm Res.2009,58(9):611-618.
[2] B.P. Singh and S.V. Gangal. Defined Allergen Extracts; Need for Efficient Diagnosis ofAllergy and Immunotherapy. Indian J Allergy Asthma Immunol.2001,15(2):67-74.
[3] Anonymous.Allergen immunotherapy:therapeutic vaccines for allergic diseases WHOposition paper. Allergy.1998,53:5-42.
[4] Anonymous. The use of standardized allergen extracts. Position statement from thecommittee on allergen standardization. American Academy of Allergy, Asthma, andImmunology(AAAAI). Allergy Clin. Immunol.1997,99:583-586.
[5] Nelson H.S. The use of standardized extracts in allergen immunotherapy. Allergy Clin.Immunol.2000,106:41-45.
[6] FAO. Report of the FAO technical consultation on food allergies. Rome1995.
[7] Daul. C. B., J. E. Morgan. and S. B. Lehrer. Hypersensitivity reactions to crustacea andmollusks. Clin. Rev. Allergy.1993,11(2):201-222.
[8] Aas K: Studies of hypersensitivity to fish. A clinical study. Int Arch Allergy Appl Immunol.1966,29:346-363.
[9] Shibaski M, Suzuki S, Tajima S, et al: Allergenicity of major component proteins ofsoybeans. Int Arch Allergy Appl Immunol.1980,61:441-448.
[10] Yunginger JW, Jones R: A review of peanut chemistry—implications for the standardizationof peanut extracts. Presented at4th Int. Paul Ehrlich Symposium on Regulatory Control andStandardization of Allergenic Extracts, Bethesda, MD,1985, October16-18.
[11] Robert F. Lemanske, Jr, and Steve L. Taylor.Standardized Extracts, Foods. CLIN REVALLERGY.1987,5:23-36.
[12] Dannaeus A, Inganas M: A follow-up study of children with food allergy. Clinical course inrelation to serum IgE-and IgG-antibody levels to milk, fish, and eggs. Clin Allergy.1981,11:533-539.
[13] Bock SA: The natural history of food sensitivity. J Allergy Clin Immunol.1982,69:173-177.
[14] Ford RPK, Taylor B: Natural history of egg hypersensitivity. Arch Dis Child.1982,57:649-652.
[15] Cherry JP, Neucere NI, Ory RL: Comparison of proteins of peanuts grown in different areas.I. Disc electrophoretic analysis of qualitative and quantitative variation. J Am Peanut ResEduc Assoc Inc.1971,3:63-74.
[16] Basha SMM: Identification of cultivar differences in seed polypeptide composition ofpeanuts (Arachis hypogaea L.) by two dimensional polyacrylamide gel electrophoresis. PlantPhysiol.1979,63:301-306.
[17] Barnett D, Baldo BH, Howden MEH: Multiplicity of allergens in peanuts. J Allergy ClinImmunol.1983,72:61-68.
[18] Arachis hypogaea L.) seed during oil roasting. J Agric Food Chem.1985,33:350-353.
[19] Perlman F: Food allergens, in Catsimpoolas N (ed): Immunological Aspects of Foods.Westport, CT, AVI Publishing Company,1977,279-316.
[20]刘锐,李冉,陈洁.我国水产品消费特征及增长潜力.农业消费展望.2011,3:53-58.
[21] FAO. Report of the FAO Technical Consultation on Food Allergies. Rome.1995.
[22] Sicherer, S.H., Munoz-Furlong, A., Sampson, H.A. Prevalence of seafood allergy in theunited states determined by a random telephone survey. J. Allergy Clin. Immunol.2004,114(1):159-165.
[23] Daul, C.B., Slattery, M., Reese, G., Lehrer, S.B. Identification of the major brown shrimp(Penaeus aztecus) allergen as the muscle protein tropomyosin. Int. Arch. Allergy Immunol.1994,105(1):49-55.
[24] Wild, L.G., Lehrer, S.B. Fish and shellfish allergy. Curr. Allergy Asthma Rep.2005,5,74-79.
[25] SAMPSON H A. Food Allergy. Part1. Immunopathogenesis and clinical disorders. J AllergyClin Immunol,1999,103(5):717-728.
[26] Daul C B,Slattery M,Reese G,et al.Identification of the major brown shrimp(Penaeusaztecus)allergen as the muscle protein tropomyosin.Int Arch Allergy ApplImmunol,1994,105:49.
[27] Hughes D A,Mills C.Food allergy:a problem on the increase. Biologist(London),2001,48(5):201-204.
[28] Musmand JJ, Daul CB, Lehrer SB. Crustacea allergy. Clin Exp Allergy.1993,23:722-32.
[29] Wang J, Sampson HA. Food anaphylaxis. Clin Exp Allergy.2007,37:65160.
[30] SAMPSON H A. Update on food allergy. Journal of Allergy and Clinical Immunology.2004,113:805-819.
[31] SAMPSON H A. Food allergy Part1: Immunopathogenesis and clinical disoders. Journal ofAllergy and Clinical Immunology.1999,103:717-728.
[32] EMMETT S E, ANGUS F J, FRY J S, et al. Perceived prevalence of peanut allergy in GreatBritain and its association with other atopic conditions and with peanut allergy in otherhouseholdmembers. Allergy.1999,54(4):380-385.
[33] MONERET-VAUTRIN D A. Epidemiologie de I allergie alimentaire et prevalence relativedes trophallergenes. Cahiers Nutrition Dietetique.2001,36(4):247-252.
[34] SCHARFER T, BOHLER E, RUHDORFER S, et al. Epidemiology of food allergy/foodintolerance in adults: associations with other manifestations of atopy. Allergy.2001,56(12):1172-1179.
[35]李振兴,林洪,李明华等.不同虾类的过敏原及其过敏原性.水产学报.2006,30(3):281-284.
[36] Goldman AS, Sellars WA, Holpern SR, et al: Milk allergy. II. Skin testing of allergic andnormal children with purified milk proteins. Pediatrics.1963,32:572-579.
[37] Lebenthal E: Cows milk protein allergy. Pediatr Clin.1975,22:827-833.
[38] Visakorpi JK, Immonen P: Intolerance to cows' milk and wheat gluten in the primarymalabsorption syndrome in infancy. Acta Pediatr Scand.1967,56:49-56
[39] Kletter B, Gery I, Freier S, et al: Immunoglobin E antibodies to milk proteins. Clin Allergy1.1971,249-255.
[40] Bleumink E, Young E: Identification of the atopic allergen in cows' milk. Int Arch AllergyAppl Immunol.1971,34:521-543.
[41] Spies JR, Stevan MA, Stein WJ, et al: The chemistry of allergens XX. New antigensgenerated by pepsin hydrolysis of bovine milk proteins, J Allergy.1970,45:208-219.
[42] Haddad ZH, Kalra V, Verma S: IgE antibodies to peptic and peptic-tryptic digests of betalactoglobulin: Significance in food hypersensitivity. Ann Allergy.1979,42:368-371.
[43] Galland A V, DoryD, Pons L. Purification of a41kD acod-allergenic protein. J Chromato B,1998,706:63-67.
[44] Van D T, Hordvik I, Endresen C. Characterization of parvalbumin, the major allergen inAlaska pollack and comparison with codfish. Mol Immuno,l.2005,42(3):345-353.
[45] Van D T, Hordvik I, Endresen C. The major allergen (parvalbumin) of codfish is encoded byat least two isotypic genes: cDNA cloning, expression and antibody binding of therecombinant allergens. Mol Immuno.2003,39(10):595-602.
[46]李振兴，林洪，李明华等.不同虾类的过敏原及其过敏原性.水产学报.2006，30(2):281-284.
[47] Chu,K.H.,Wong,S.H.,and Leung,P.S.Tropomyosin is the major mollusk allergen:reversetranscriptase polymerase chain reaction, expression and IgE reactivity. Mar Biotechnol.2000,2(5):499~509.
[48]毛露甜,范红英,张在军,黄婉慧,杨华胜,周立斌.虾类和蟹类的过敏原组分分析.细胞与分子兔疫学杂志.2008,24(6):642-644.
[49]毛露甜,黄婉慧,吴小明,范红英,张在军.虾类和蟹类过敏原的交叉反应性研究.中国免疫学杂志.2008,24:748-752.
[50] Platts-Mills T.A.E., Chapman M.D. Allergen standardization J. Allergy Clin. Immunol.1991,87:621-625.
[51] Frankland A.W. The characterization of allergens. Clin. Exp. Allergy.1996,26:865-867.
[52] Anonymous. Allergen nomenclature. WHO/IUIS Allergen nomenclature subcommittee WorldHealth Organization, Geneva, Switzerland. Clin. Exp. Allergy.1995,25:21-37.
[53] Robret E.Reisman, M.D., Review of Procedures Used in the Practice of Allergy,1981,57(7):549-558.
[54]邓鹏，孙守义，王守经，张奇志，食品过敏原的安全管理，中国食物与营养，2008,12-14.
[1] Jona,R., Fronda,A., Comparative histochemical analysis of cell wall polysaccharides byenzymatic and chemical extractions of two fruits, Biotech. Histochem.1997,72:22-28.
[2] Smith, P.K., Krohn R.I., Hermanson, G.TT.,Mallia, A.K., Gartner, F. H., Provenzano,M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. Measurement of protein usingbicinchoninic acid. Anal. Biochem.1985,150(1):76-85.
[3]张轶群,李振兴,林洪,等.果糖和木糖在美拉德反应中对虾类过敏原活性影响的研究.食品科学,2009,30(29):11-14.
[4] Motoyama K., Suma Y., Ishizaki S., Nagashima Y., Shiomi K. Molecular cloning oftropomyosins identified as allergens in six species of Crustaceans. Journal of Agriculturaland Food Chemistry,2007,55(3):985-991.
[5] Steinhoff, M.; Fischer, M.; Paschke-Kratzin, A. Comparison of extraction conditions for milkand hen's egg allergens. Food Additives&Contaminants: Part A.2011,28,(4):373-383.
[6]刘一璇,林洪,吴丽莎,等.加工方式对刀额新对虾主要过敏原免疫原性的影响.水产学报.2011,35(6):948-953.
[7] Ruegg. UT, Rudinger. J.“Reductive cleavage of cystine disulfides withtributylphosphine”. Methods Enzymol.1977,47:111-116.
[8] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V., Identification oftropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes. J.Immunol.1993,151(10):5354-5363.
[9] Motoyama, K., Suma, Y., Ishizaki, S., Nagashima, Y., Shiomi, K. Molecular cloning oftropomyosins identified as allergens in six species of crustaceans. J. Agric. Food Chem.2007,55(3):985-991.
[10] D. Wessel, U.I. Flügge. A method for the quantitative recovery of protein in dilute solution inthe presence of detergents and lipids. Analytical Biochemistry.1984,138(1):141-143.
[1] FAO/WHO. Evaluation of allergenicity of genetically modified foods, in Report of a JointFAO/WHO Expert Consultation of Allergenicity of Foods Derived from Biotechnology.World Health Organization, Geneva.2001,22-25.
[2] Lopata, A.L., Lehrer, S.B. New insights into seafood allergy. Curr Opin Allergy ClinImmunol.2009,9:270-277.
[3] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V. Identification oftropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes. J.Immunol.1993,151(10):5354-5363.
[4] Daul, C.B., Slattery, M., Reese, G., Lehrer, S.B. Identification of the major brown shrimp(Penaeus aztecus) allergen as the muscle protein tropomyosin. Int. Arch. Allergy Immunol.1994,105(1):49-55.
[5] Leung, P.S., Chu, K.H., Chow, W.K., Ansari, A., Bandea, C.I., Kwan, H.S., Nagy, S.M.,Gershwin, M.E. Cloning, expression, and primary structure of Metapenaeus ensistropomyosin, the major heat-stable shrimp allergen. J. Allergy Clin. Immunol.1994,94(5):882-890.
[6] Leung, P.S.C., Chen, Y.S., Mykles, D.L., Chow, W.K., Li, C.P., Chu, K.H. Molecularidentification of the lobster muscle protein tropomyosin, as a seafood allergen. Mol. Mar.Biol. Biotechnol.1998,7:12-20.
[7] Bauermeister, K., Wangorsch, A., Perono Garoffo, L., Reuter, A., Conti, A., Falk, S., Taylor,S.L., Vieths, S., Holzhauser, T., Ballmer-Weber, B.K., Reese, G. Novel Crustacean AllergensIdentified in North Sea Shrimp Crangon and other Crustacean Species-SarcoplasmicCalcium-binding Protein, Troponin C, Troponin I, Triosephosphate Isomerase, and MyosinLight Chain.2008, http://www.ncbi.nlm.nih.gov/protein/238477329.
[8] Shiomi, K., Sato, Y., Hamamoto, S., Mita, H., Shimakura, K. Sarcoplasmic calciumbindingprotein: identification as a new allergen of the black tiger shrimp Penaeus monodon. Int. Arch.Allergy Immunol.2008,146:91-98.
[9] Ayuso, R., Grishina, G., Ibáňez, M.D., Blanco, C., Carrillo, T., Bencharitiwong, R., Sánchez,S., Nowak-Wegrzyn, A., Sampson, H.A. Sarcoplasmic calcium-binding protein is anEF-hand-type protein identified as a new shrimp allergen. J.Allergy Clin. Immunol.2009,124:114-120.
[10] Rosal′a Ayuso, MD, PhD, Galina Grishina, MS, Ludmilla Bardina, MS, Teresa Carrillo, MD,PhD, Carlos Blanco, MD, PhD, María Dolores Ibáňez, MD, PhD, Hugh A. Sampson, MDand Kirsten Beyer, MD. Myosin light chain is a novel shrimp allergen, Lit v3. AmericanAcademy of Allergy, Asthma&Immunology.2008,122:795-802.
[11] Ayuso, R., Grishina, G., Bardina, L., Carrillo, T., Blanco, C., Ibaňez, M.D., et al Myosin lightchain is a novel shrimp allergen, lit v3. J. Allergy Clin. Immunol.2008,122(4)：795-802.
[12] Yu, C.J., Lin, Y.F., Chiang, B.L., Chow, L.P. Proteomics and immunological analysis ofanovel shrimp allergen, Pen m2. J. Immunol.2003,170:445-453.
[13] Gar a-Orozco, K.D., Aispuro-Hern′andez, E., Yepiz-Plascencia G, Calde on-de-la-Barca,A.M., Sotelo-Mundo, R.R. Molecular characterization of arginine kinase, an allergen fromthe shrimp Litopenaeus vannamei. Int. Arch. Allergy Immunol.2007,144:23-28.
[14] Smith, P.K., Krohn R.I., Hermanson, G.TT.,Mallia, A.K., Gartner, F. H., Provenzano,M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. Measurement of protein usingbicinchoninic acid. Anal. Biochem.1985,150(1):76-85.
[15]张轶群,李振兴,林洪等.果糖和木糖在美拉德反应中对虾类过敏原活性影响的研究.食品科学.2009,30(29):11-14.
[16] Leung, P.S.C., Chen, Y.S., Gershwin, M.E., Wong, S.H., Kwan, H.S., Chu, K.H.Identification and molecular characterization of Charybdis feriatus tropomyosin, the majorcrab allergen. J. Allergy Clin. Immunol.1998,102:847-852.
[17] Shanti, K.N., Martin, B.M., Nagpal, S., Metcalfe, D.D., Rao, P.V. Identification oftropomyosin as the major shrimp allergen and characterization of its IgE binding epitopes. J.Immunol.1993,151(10):5354-5363.
[18] Motoyama, K., Suma, Y., Ishizaki, S., Nagashima, Y., Shiomi, K. Molecular cloning oftropomyosins identified as allergens in six species of crustaceans. J. Agric. Food Chem.2007,55(3):985-991.
[19] Bredehorst, R., David, K. What establishes a protein as an allergen? J. Chromatogr. B.Biomed Sci Appl.2001,756,33-40.
[20] Leung, P. S. C., Chow, W. K., Duffey, S., Kwan, H. S., Gershwin, M. E., Chu, K. H. IgE
reactivity against a cross-reactive allergen in crustacea and mollusca: evidence for
tropomyosin as the common allergen. J. Allergy Clin. Immunol.1996,98:954-961.
[1] Surapon Piboonpocanun, Siribangon Boonchoo, Wipada Pariyaprasert, NualanongVisitsunthorn and Orathai Jirapongsananuruk. Determination of storage conditions for shrimpextracts: analysis of specific IgE-allergen profiles. Asian Pacific Journal of Allergy andImmunology.2010,28:47-52.
[2] Jirapongsananuruk O, Sripramong C, Pacharn P, et al. Specific allergy to Penaeus monodon(seawater shrimp) or Macrobrachium rosenbergii (freshwater shrimp) in shrimp-allergicchildren. Clin Exp Allergy.2008,38:103847.
[3] Carnes J, Ferrer A, Huertas AJ, Andreu C, Larramendi CH, Fernandez-Caldas E. The use ofraw or boiled crustacean extracts for the diagnosis of seafood allergic individuals. Ann
Allergy Asthma Immunol.2007,98:34954.
[1]赵俊芳,王学谦.食物过敏原检测及其应用前景[J].中华检验医学杂志.2007,30(8):948-950.
[2]贺娟,张玉,肖会.阿罗格脱敏治疗支气管哮喘的疗效观察及护理[J].现代中西医结合杂志.2009,18(19):2336-2337.
[3]匡嘉丽,方向佳,吴军等.阿罗格长效变应原脱敏治疗变应性鼻炎的疗效观察[J].海南医学院学报.2009,15(8):883-886.
[4]张春林.阿罗格标准化变应原特异性免疫治疗对儿童过敏性哮喘的持续影响[J].药物与临床.2009,47(12):89-90.
[5] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685.
[6] MOTOYAMA K., SUMA Y., ISHIZAKI S. et al. Molecular cloning of tropomyosinsidentified as allergens in six species of Crustaceans. Journal of Agricultural and FoodChemistry.2007,55(3):985-991.
[7] WATSON P.F. The causes of reduced fertility with cryopreserved semen. AnimalReproduction Science.2000,60-61:481-492.
[8]杨继东.冻融对血浆总蛋白含量的影响[J].现代医药卫生.2004,20(3):1271.
[9] XIUFANG XIA, BAOHUA KONG, QIAN LIU, et al. Physicochemical change and proteinoxidation in porcine longissimus dorsi as inuenced by different freeze-thaw cycles. MeatScience.2009,83:239-245.
[10] SADETTIN TURHAN, N. SULE USTUN, INCI BANK. Effect of freeze-thaw cycles ontotal and heme iron contents of bonit (Sardasarda) and bluesh (Pomatomus saltator) fillets.Journal of Food Composition and Analysis.2006,19:384-387.
[11] N.S.PRATHALINGAM, S.G.REVELL, S.MIRCZUK, et al. Impact of antifreeze proteins andantifreeze glycoproteins on bovine sperm during freeze-thaw. Theriogenology.2006,66:1894-1900.
[12] STROM C S, LIU XY, JIA Z. Antifreeze protein-induced morphological modificationmechanisms linked to ice binding surface. Biological Chemistry.2004,279(31):32407-32417.
[13] SHANN-TZONG JIANG, DER-CHYAN HWANG, CHING-SAN CHEN. Effect of storagetemperature on the formation of disulfides and denaturation of milkfish actomyosin. Foodscience.1988,53(5):1333-1335.
[14]曾名勇,黄海,李八方.鳙肌肉蛋白质生化特性在冻藏过程中的变化[J].水产学报,2003,27(5):480-485.
[15]曾名勇,黄海,李八方.不同冻藏温度对鲈鱼肌肉蛋白质生化特性的影响[J].青岛海洋大学学报,2003,33(33):525-530.
[1]吴丽莎，李振兴，刘一璇，陈瑜，林洪.冻融循环过程中虾过敏原的免疫原性变化.水产学报.2011,35(5):137-141
[2] Wakamatsu T, Sato Y. Determination of unfreezable water in sucrose, sodium chlorideandprotein solutions by differential scanning calorimeter. Nippon Nogeikagaku Kaishi,1979,53：415-420.
[3] Wakamatsu T，Saito Y. Further study on the effect of sucrose and sodium chloride on gelatinand unfreezable water of egg yolk during freezing. Nippon Nogeikagaku Kaishi,1980,54:951-957.
[4] Matsumotom I,Ooizumi T,Arai K.Protective effect of sugar on freeze-denaturation of carpmyofibrillar protein. Nippon Suisan Gakkaish,1985,51:833-839.
[5]曲楠，曾名湧，董士远.海藻糖对罗非鱼糜蛋白抗冻作用的研究[J].肉类研究,2010,(1):79-82.
[6]薛勇,薛长湖,李兆杰,等.海藻糖对冻藏过程中鳙肌原纤维蛋白冷冻变性的影响[J].2006,13:637-641.
[7] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685.
[8]李振兴,林洪,李明华等.不同虾类的过敏原及其过敏原性[J].水产学报.2006,30(3):281-284.
[9] Kauzmann W. Some factors in the interpretation of protein denaturation. Protein Chemistry，1959,14:1-63.
[10] Hanafusa N.Biological macromolecule and water [C]//Japanese Society of Fisheries Science.Food and Water.Koseisha Koseikaku，Tokyo.1973:9-24.
[11]严宏忠.鱼肉蛋白质的冷冻变性及防止方法[J].中国水产,1995,(3):37-38.
[12]张娅楠,赵利,刘华,袁美兰,邓丹雯.水产品的冷冻变性及鱼糜抗冻剂研究进展.[J].河南工业大学学报(自然科学版),2011,32(6):88-92.
[1] Taylor, S.L., Lehrer, S.B. Principles and characteristics of food allergens. Crit. Rev. Food Sci.Nutr.1996,36(S):91-118.
[2] Huang, Y.Y., Liu, G.M., Cai, Q.F., Weng, W.Y., Maleki, S.J., Su, W.J., Cao, M.J. Stabilityof major allergen tropomyosin and other food proteins of mud crab (Scylla serrata) by invitro gastrointestinal digestion. Food Chem. Toxicol.2010,48,1196-1201.
[3] Astwood J D，Leach J N，Fuchs R L.Stability of food allergens to digestion in vitro. NatBiotechnol.1996,14(10):1269-1273.
[4] Kenna J G，Evans R M.Digestibility of proteins in simulated gastric fluid.Toxicologist.2000,54:141.
[5] Fu T J.Digestion stability as a criterion for protein allergenicity assessment. Ann N Y AcadSci.2002,964:99-110.
[6] Thomas, K., Aalbers, M., Bannon, G.A., Bartels, M., et al. A multi-laboratory evaluation of acommon in vitro pepsin digestion assay protocol used in assessing the safety of novelproteins. Regul. Toxicol. Pharmacol.2004,39(2):87-98
[7] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head ofbacteriophage T4. Nature.1970,227:680-685
[8] Liu, G.M., Huang, Y.Y., Cai, Q.F., Weng, W.Y., Su, W.J., Cao, M.J. Comparative study of invitro digestibility of major allergen, tropomyosin and other proteins between Grass prawn(Penaeus monodon) and Pacific white shrimp (Litopenaeus vannamei). J. Sci. Food Agric.2011,91:163-170.
[9] Fu T J，Abbott U R，Hatzos C. Digestibility of food allergens and nonallergenic proteins insimulated gastric fluid and simulated intestinal fluid—A comparative study. J Agric FoodChem.2002,50:7154-7160.
[1] Passante E,Ehrhardt C,Sheridan H,et al.RBL-2H3cells arean imprecise model for mast cellmediator release. Inflamm Res.2009,58(9):611-618.
[2] Eccleston E，Leonard BJ，Lowe JS，et al. Basophilic leukaemia in the albino rat and ademonstration of the basopoietin. Nat New Biol.1973,44:73-76.
[3] Hoffmann, A.Jamin, K. Foetiseh, et al. Determination of the allergenic activity of birehpollen and apple prick test solutions by measurement of β-hexosaminidase release fromRBL-2H3cells: Comparison with classieal methods in allergen standardization. Allergy.1999,54:446-454.
[4] Hisashi M,Sachiko S,Toshio M,et al.Bioactive constituents from Chinese naturalmedicines—Inhibitors of antigen-induced degranulation in RBL-2H3cells from the seeds ofPsoralea corylifolia. Chem Pharm Bull.2007,55:106-110.
[5] S.Kaul, D.Luttkopf, B.Kastner, L.Vogel, G.H ltz, S.Vieths and A.Hoffmann. Mediator releaseassays based on human or murine immunoglobulin E in allergen standardization. Clinicaland Experimental Allergy.2007,37,141-150.
[6] Ward MD, Selgrade MK. Animal models for protein respiratory sensitizers. Methods.2007,(41):80-90.
[7] Susanne Kaul, Stephan Scheurer, Nicole Danz, Simone Schicktanz, Stefan Vieths,Andreas Hoffmann. Monoclonal IgE antibodies against birch pollen allergens: Noveltools for biological characterization and standardization of allergens. Allergy ClinImmunol.2003,111(6):1262-1268.
[8] Sawanee Kraithep, Kwunchit Oungbho, Supinya Tewtrakul. Anti-allergic activity ofThai medicinal plants used in longevity formulation. Songklanakarin J. Sci. Technol.2008,30(5):621-625.
[9] Tomohiro Itoh, Yasunori Fujita, Mikako Ito, Akio Masuda, Kinji Ohno, MasatoshiIchihara, Toshio Kojima, Yoshinori Nozawa, Masafumi Ito. Molecular hydrogensuppresses FcεR I-mediated signal transduction and prevents degranulation of mastcells. Biochemical and Biophysical Research Communications.2009,389(4):651-656.
[10] Eun-Kyung PARK, Min-Kyung CHOO, Eun-Jin KIM, Myung Joo HAN, Dong-HyunKIM. Antiallergic activity of ginsenoside Rh2. Biological and PharmaceuticalBulletin.2003,26(11):1581-1584.
[11] LU Y M，LI L，LUO W T，et al. A study on the change of calcium signal in mast cellactivated by antigen.Chin J Tubere Respir Dis.2000,23(4):243-246.
[12] Passante E，Frankish N. The RBL-2H3cell line: its provenance and suitability as amodel for the mast cell. Inflamm Res.2009,58(11):737-745.
[13] R.J. Dearman，R.A. Skinner，N. Deakin，et al. Evaluation of an in vitro method for themeasurement of specific IgE antibody responses: the rat basophilic leukemia (RBL) cellassay. Toxicology.2005,206:195-205.
[14] Passante E，Ehrhardt C，Sheridan H，et al. RBL-2H3cells are an imprecise model for mastcell mediator release. Inflamm Res.2009,58(9):611-618.