本文選題：戊二醛聚合方法 + 血紅蛋白聚合產(chǎn)物��； 參考：《西北大學(xué)》2009年碩士論文

【摘要】： 血紅蛋白類氧載體(Hemoglobin-based oxygen carriers,HBOCs)是一類通過(guò)血紅蛋白分子表面化學(xué)修飾或分子間交聯(lián)而形成分子量較大的血紅蛋白制品,原料血紅蛋白主要來(lái)源于人、豬、牛,常用戊二醛作為交聯(lián)劑,具有無(wú)病原微生物污染、制劑組成相對(duì)簡(jiǎn)單、可長(zhǎng)期體外保存、使用時(shí)不需要交叉配型等優(yōu)點(diǎn)。但此類制劑也存在潛在的問(wèn)題,即異源蛋白和蛋白質(zhì)修飾/聚合反應(yīng)可能使得此類產(chǎn)品刺激或抑制機(jī)體免疫系統(tǒng)功能,從而影響機(jī)體體液免疫和細(xì)胞免疫,對(duì)機(jī)體產(chǎn)生免疫毒性作用。免疫原性是免疫毒性評(píng)價(jià)的重要組成部分,目前已成為研究重點(diǎn)。本文主要采用抗體生成反應(yīng)方法,初步研究了戊二醛聚合反應(yīng)前后豬、牛血紅蛋白的免疫原性所發(fā)生的變化,以研究聚合反應(yīng)對(duì)此兩類蛋白免疫原性的影響。 實(shí)驗(yàn)結(jié)果顯示,首先豬、牛血紅蛋白刺激機(jī)體生成抗體的作用都很弱,但經(jīng)過(guò)戊二醛聚合后體液免疫刺激作用明顯增強(qiáng);其次,抗牛血紅蛋白抗體不識(shí)別聚合牛血紅蛋白,抗分子內(nèi)聚合牛血紅蛋白抗體也不識(shí)別天然牛血紅蛋白,而分子間聚合牛血紅蛋白抗體卻可以識(shí)別天然的牛血紅蛋白;最后,抗豬血紅蛋白抗體可識(shí)別聚合豬血紅蛋白,抗聚合豬血紅蛋白抗體也可識(shí)別天然豬血紅蛋白。 可見(jiàn),豬、牛血紅蛋白免疫原性很低,但是戊二醛聚合反應(yīng)改變了蛋白質(zhì)的結(jié)構(gòu),使其免疫刺激作用顯著增強(qiáng),提高了豬、牛血紅蛋白刺激機(jī)體生成抗體的能力。另外,不同蛋白質(zhì)空間結(jié)構(gòu)的改變對(duì)免疫原性產(chǎn)生影響的機(jī)理可能不同。戊二醛分子內(nèi)聚合反應(yīng)可能沒(méi)有改變豬血紅蛋白抗原決定簇的結(jié)構(gòu),但卻改變了牛血紅蛋白抗原決定簇的結(jié)構(gòu),而分子間聚合對(duì)牛血紅蛋白抗原決定簇沒(méi)有影響。戊二醛聚合反應(yīng)可造成此兩類蛋白免疫學(xué)性質(zhì)發(fā)生不同改變的事實(shí)說(shuō)明,雖然不同種血紅蛋白的氨基酸序列具有很高的相似性,但聚合反應(yīng)所產(chǎn)生的免疫學(xué)特性的影響可能完全不同。這種影響與聚合反應(yīng)所修飾的蛋白質(zhì)的部位及其相鄰空間結(jié)構(gòu)的特點(diǎn)有密切關(guān)系。同種化學(xué)修飾方法制備的不同來(lái)源血紅蛋白的聚合產(chǎn)物可具有不同的免疫學(xué)刺激特性,而不同種化學(xué)修飾方法制備的相同來(lái)源血紅蛋白產(chǎn)物也具有不同的免疫學(xué)特性。 因此,必須分別研究不同種HBOCs產(chǎn)品的免疫學(xué)特性,不能僅根據(jù)某一HBOCs產(chǎn)品分子的免疫學(xué)特性推斷用相同制備方法獲得的不同來(lái)源的血紅蛋白產(chǎn)品或用不同制備方法獲得的相同來(lái)源的血紅蛋白產(chǎn)品的免疫學(xué)特征。
[Abstract]:Hemoglobin-based oxygen carriers (HBOCs) is a kind of hemoglobin products with high molecular weight, which is formed by chemical modification or intermolecular crosslinking of hemoglobin molecules. The raw materials of hemoglobin-based hemoglobin are mainly from human, pig and cow.The common glutaraldehyde as crosslinking agent has the advantages of no contamination of pathogenic microorganisms, relatively simple composition, long-term preservation in vitro and no cross-matching when used.But there are also potential problems with such preparations, that is, heterologous proteins and protein modification / polymerization may cause such products to stimulate or inhibit the immune system of the body, thereby affecting the humoral and cellular immunity of the body.Immune toxicity to the body.Immunogenicity is an important part of immunotoxicity evaluation, and has become the focus of research.In this paper, the immunogenicity of porcine and bovine hemoglobin before and after glutaraldehyde polymerization was studied by antibody forming reaction, and the effect of polymerization on the immunogenicity of these two proteins was studied.The results showed that, first of all, pig and bovine hemoglobin stimulated the body to produce antibody very weak, but after glutaraldehyde polymerization, the humoral immune stimulation effect was obviously enhanced; secondly, the anti-bovine hemoglobin antibody did not recognize polymerized bovine hemoglobin.Anti-intramolecular polymerized bovine hemoglobin antibodies also do not recognize natural bovine hemoglobin, whereas intermolecular polymerized bovine hemoglobin antibodies can recognize natural bovine hemoglobin; finally, anti-porcine hemoglobin antibodies can recognize polymerized porcine hemoglobin.Anti-polymeric porcine hemoglobin antibodies can also recognize natural porcine hemoglobin.It can be seen that the immunogenicity of pig and bovine hemoglobin is very low, but glutaraldehyde polymerization has changed the structure of protein, enhanced its immunostimulatory effect, and improved the ability of pig and bovine hemoglobin to stimulate the body to produce antibody.In addition, the effects of different protein spatial structure on immunogenicity may be different.The intramolecular polymerization of glutaraldehyde may not change the structure of porcine hemoglobin antigen determinant, but it changes the structure of bovine hemoglobin antigen determinant, while intermolecular polymerization has no effect on bovine hemoglobin antigen determinant.The fact that glutaraldehyde polymerization can result in different immunological properties of these two proteins suggests that although the amino acid sequences of different hemoglobin are highly similar,But the effects of polymerization on immunological properties may be completely different.This effect is closely related to the site of the modified protein and the characteristics of its adjacent spatial structure.The polymerized products from different sources of hemoglobin prepared by the same chemical modification method may have different immunological stimulation properties, and the same source hemoglobin products prepared by different chemical modification methods also have different immunological characteristics.Therefore, it is necessary to study the immunological characteristics of different HBOCs products,The immunological characteristics of hemoglobin from different sources obtained by the same preparation method or from the same source by different preparation methods should not be inferred from the immunological properties of a certain HBOCs product molecule.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 牛秀杰;禽網(wǎng)狀內(nèi)皮組織增殖癥病毒三種檢測(cè)方法的建立[D];東北農(nóng)業(yè)大學(xué);2013年

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本文編號(hào)：1769152