【摘要】：人凝血因子Ⅷ、人凝血酶原復(fù)合物、人凝血因子Ⅸ等因子類(lèi)血液制品,是以健康人血漿為原料,經(jīng)分離純化和病毒滅活制成,在臨床上主要用于各種凝血因子缺乏的治療,這些因子類(lèi)血液制品的凝血因子效價(jià)是藥品有效性的指標(biāo),凝血因子效價(jià)的測(cè)定也是藥品質(zhì)量檢測(cè)中的關(guān)鍵檢測(cè)項(xiàng)目。血液制品生產(chǎn)企業(yè)及各級(jí)檢測(cè)機(jī)構(gòu)一般采用全自動(dòng)血凝儀并按照儀器使用說(shuō)明來(lái)進(jìn)行因子效價(jià)的檢測(cè),然而在不同檢測(cè)方法結(jié)果比對(duì)中,發(fā)現(xiàn)全自動(dòng)血凝儀檢測(cè)結(jié)果與中國(guó)藥典中一期法的檢測(cè)結(jié)果在部分因子效價(jià)檢測(cè)中存在檢測(cè)差異,差異可能是樣品稀釋中使用的稀釋液的蛋白含量不同引起的,故本課題主要研究因子類(lèi)血液制品中蛋白質(zhì)含量對(duì)效價(jià)檢測(cè)結(jié)果影響。因子類(lèi)血液制品是血液中凝血因子的高純度濃縮物,在進(jìn)行效價(jià)測(cè)定時(shí)需要對(duì)樣品進(jìn)行稀釋,而稀釋液中蛋白含量的不同對(duì)其檢測(cè)結(jié)果有著影響,國(guó)內(nèi)因子類(lèi)血液制品的生產(chǎn)企業(yè)極少,對(duì)因子產(chǎn)品效價(jià)檢測(cè)方法的研究也很少,并主要對(duì)因子效價(jià)檢測(cè)的操作方式、實(shí)驗(yàn)條件、輔料影響等進(jìn)行了研究,本課題以尋找實(shí)際工作中不同檢測(cè)方法間結(jié)果差異的原因?yàn)槌霭l(fā)點(diǎn),確立了不同因子類(lèi)產(chǎn)品的自身性質(zhì)對(duì)效價(jià)檢測(cè)結(jié)果的影響以及不同種類(lèi)稀釋液中蛋白含量對(duì)效價(jià)檢測(cè)結(jié)果的影響,最后通過(guò)方法學(xué)驗(yàn)證確立了適用于因子效價(jià)檢測(cè)的稀釋方法,從而完善了因子類(lèi)血液制品效價(jià)檢測(cè)的方法。全自動(dòng)血凝儀主要設(shè)計(jì)用于臨床血漿樣本的檢測(cè),故在建立因子類(lèi)血液制品的全自動(dòng)血凝儀法效價(jià)檢測(cè)時(shí),需要消除產(chǎn)品本身性質(zhì)對(duì)檢測(cè)結(jié)果的影響,并對(duì)建立的檢測(cè)方法進(jìn)行方法學(xué)驗(yàn)證。本課題通過(guò)蛋白含量對(duì)因子效價(jià)檢測(cè)影響的研究建立起的全自動(dòng)血凝儀因子效價(jià)檢測(cè)方法,消除了產(chǎn)品本身性質(zhì)的影響,可以準(zhǔn)確進(jìn)行因子效價(jià)的測(cè)定,方法的建立不但可以準(zhǔn)確控制因子類(lèi)血液制品的質(zhì)量,保證生產(chǎn)企業(yè)持續(xù)穩(wěn)定生產(chǎn)出質(zhì)量均一的藥品,而且可以減少因子類(lèi)產(chǎn)品在臨床使用中的風(fēng)險(xiǎn)。本課題從工作過(guò)程中檢測(cè)結(jié)果的差異入手,對(duì)蛋白質(zhì)含量在因子效價(jià)檢測(cè)中的影響進(jìn)行了研究,并以國(guó)外同類(lèi)產(chǎn)品進(jìn)行檢測(cè)比對(duì),最后確立了全自動(dòng)血凝儀因子效價(jià)檢測(cè)的方法并對(duì)方法進(jìn)行了驗(yàn)證。具體研究?jī)?nèi)容如下：(1)對(duì)同一批人凝血因子Ⅷ、人凝血酶原復(fù)合物樣品選用不同的稀釋液進(jìn)行全自動(dòng)血凝儀法的效價(jià)檢測(cè),以確定不同稀釋液稀釋檢測(cè)結(jié)果的差別,并對(duì)連續(xù)生產(chǎn)的人凝血因子Ⅷ、人凝血酶原復(fù)合物進(jìn)行不同稀釋液的效價(jià)檢測(cè),確定不同稀釋液稀釋檢測(cè)結(jié)果的差別。(2)對(duì)同一批人凝血因子Ⅷ、人凝血酶原復(fù)合物用含不同蛋白質(zhì)濃度的稀釋液或凝血因子缺乏血漿進(jìn)行稀釋,確定蛋白質(zhì)濃度對(duì)因子效價(jià)檢測(cè)的影響,并對(duì)2批國(guó)外生產(chǎn)的同類(lèi)產(chǎn)品進(jìn)行同樣的試驗(yàn),確定出適用于因子效價(jià)檢測(cè)的稀釋方法。(3)建立起人凝血因子Ⅷ、人凝血酶原復(fù)合物的全自動(dòng)血凝儀因子效價(jià)檢測(cè)方法并進(jìn)行方法驗(yàn)證,選擇適用于因子效價(jià)檢測(cè)的稀釋方法和稀釋液,對(duì)方法的線性關(guān)系、準(zhǔn)確度、精密度、日間精密度、專(zhuān)屬性等進(jìn)行了方法學(xué)驗(yàn)證,確認(rèn)所建立方法適用于因子類(lèi)血液制品效價(jià)的檢測(cè)。
[Abstract]:human blood coagulation factor VIII, human prothrombin complex, human blood coagulation factor IX and other factor-like blood product are prepared from human plasma as raw material, separated and purified and virus-inactivated, and is mainly used for treating various blood coagulation factor deficiency in clinic, The coagulation factor titer of these factor-based blood products is an index of the effectiveness of the drug, and the determination of the potency of the blood coagulation factor is also a key detection item in the quality detection of the drug. The blood product production enterprise and the detection mechanism at all levels generally adopt a full-automatic blood coagulation instrument and carry out the detection of the factor titer according to the instructions of the instrument, It is found that the detection result of the full-automatic blood-coagulation instrument and the detection result of the first-phase method in the Chinese pharmacopoeia are different from the detection result of the first-phase method in the Chinese pharmacopoeia, and the difference can be caused by the difference in the protein content of the diluent used in the sample dilution, Therefore, the content of protein in the main research factor blood product of the subject is affected by the result of the potency test. The factor-based blood product is a high-purity concentrate of the blood coagulation factor in the blood, The research on the method for detecting the potency of the factor product is very small, and the operation mode, the experimental conditions and the influence of the auxiliary materials of the factor potency test are also studied. The main cause of the difference between the different detection methods in the actual work is the starting point. The effect of the self-nature of the different factor products on the potency test result and the effect of the protein content on the titer detection result in different kinds of diluent are established, and finally, the dilution method applicable to the detection of the factor titer is established through the method verification, So as to improve the method for detecting the potency of the factor-like blood product. The full-automatic blood-coagulation instrument is mainly designed for the detection of clinical plasma samples, so that the effect of the nature of the product on the detection result needs to be eliminated when the full-automatic blood-coagulation instrument of the factor-based blood product is established, and the established detection method is validated. The method for detecting the potency of the full-automatic blood-coagulation instrument established by the research on the influence of the protein content on the titer detection of the factor eliminates the influence of the properties of the product, and can accurately carry out the measurement of the factor titer, The establishment of the method not only can accurately control the quality of the factor-based blood products, ensure the continuous and stable production of the drugs with uniform quality, but also can reduce the risk of the factor products in clinical use. In this paper, the influence of the protein content in the detection of the factor titer was studied by the difference of the detection results in the working process, and the method for detecting the potency of the full-automatic blood-coagulation instrument was established and the method was validated. The specific research contents are as follows: (1) the same batch of human blood coagulation factor VIII and human prothrombin complex samples are selected to carry out the full-automatic blood-coagulation instrument titer detection to determine the difference of the dilution detection result of different dilutions, and the human blood coagulation factor VIII is continuously produced, The value of different dilutions of human prothrombin complex was tested to determine the difference in the dilution test results for different dilutions. (2) the same batch of human blood coagulation factor VIII and human prothrombin complex are diluted with a diluent or a blood coagulation factor containing different protein concentration to determine the influence of the protein concentration on the detection of the factor titer, and the same test is carried out on the similar products produced in the two batches, A dilution method suitable for the detection of a factor titer is determined. and (3) establishing a full-automatic blood coagulation instrument factor titer detection method for the human blood coagulation factor VIII and the human prothrombin complex and carrying out a method verification, selecting a dilution method and a diluent suitable for the detection of the factor titer, the linear relationship, the accuracy, the precision and the day precision of the method, The specificity and the like are verified by the methodology, and the established method is applicable to the detection of the potency of a factor-based blood product.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R977.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 李青;劉曉;張t，

本文編號(hào)：2488489