發(fā)布時(shí)間：2018-05-06 02:25

  本文選題：阿爾茨海默病 + 人源化抗體　； 參考：《基礎(chǔ)醫(yī)學(xué)與臨床》2010年08期

【摘要】：目的將前期構(gòu)建的抗Aβ人-鼠嵌合抗體進(jìn)行可變區(qū)人源化改造,為抗Aβ抗體在臨床人體中應(yīng)用奠定基礎(chǔ)。方法采用模板替換法對已構(gòu)建的人-鼠嵌合抗體基因中的重、輕鏈可變區(qū)框架區(qū)進(jìn)行人源化改造,構(gòu)建抗Aβ人源化抗體的真核表達(dá)載體。用脂質(zhì)體法將重、輕鏈共轉(zhuǎn)染CHO細(xì)胞進(jìn)行表達(dá),采用ELISA法檢測表達(dá)產(chǎn)物效價(jià)、表達(dá)量;同時(shí)用SDS-PAGE及Western blot檢測表達(dá)產(chǎn)物分子質(zhì)量;免疫組化法鑒定其生物學(xué)活性。結(jié)果重組改造后的抗Aβ人源化抗體基因重鏈約為1 500 bp,輕鏈約為750 bp,與預(yù)期一致。轉(zhuǎn)染CHO細(xì)胞后獲得表達(dá),表達(dá)量為1.11 mg/L,抗體重鏈相對分子質(zhì)量約為51 ku,輕鏈約為25 ku。ELISA結(jié)果顯示能與Aβ特異性結(jié)合(細(xì)胞培養(yǎng)上清A值:2.24),與改造前的嵌合抗體比較效價(jià)基本相同�？贵w人源化檢測顯示,只能被羊抗人抗體識(shí)別,不能被羊抗鼠抗體識(shí)別。免疫組化顯示表達(dá)產(chǎn)物有結(jié)合Aβ抗原的活性。結(jié)論將抗Aβ人-鼠嵌合抗體進(jìn)行可變區(qū)人源化改造后,基本保持了原嵌合抗體的生物學(xué)特性,為其今后應(yīng)用于臨床提供了可能性。
[Abstract]:Objective to humanize the previously constructed anti A 尾 human mouse chimeric antibody in order to lay a foundation for the clinical application of anti A 尾 antibody. Methods template replacement method was used to humanize the heavy and light chain variable frame region of the constructed human / mouse chimeric antibody gene, and the eukaryotic expression vector of anti A 尾 humanized antibody was constructed. CHO cells were cotransfected with heavy and light chains by liposome method. The titer and quantity of the expression products were detected by ELISA method. The molecular weight of the expressed products was detected by SDS-PAGE and Western blot. The biological activity of the expressed products was identified by immunohistochemical method. Results the heavy chain of anti-A 尾 humanized antibody gene was about 1 500 BP, and the light chain was about 750 BP, which was consistent with the expectation. After transfection of CHO cells, the expression amount was 1.11 mg / L, the relative molecular weight of antibody heavy chain was about 51 ku. and the light chain was about 25 ku.ELISA. The results showed that the cell culture supernatant A: 2.24 could bind to A 尾 specifically. The titer of chimeric antibody was basically the same as that of chimeric antibody before modification. Antibody humanization test showed that only sheep anti-human antibody can be recognized, not sheep anti-mouse antibody. Immunohistochemistry showed that the expressed product had the activity of binding A 尾 antigen. Conclusion after the humanization of anti-A 尾 human-mouse chimeric antibody, the biological characteristics of the original chimeric antibody were basically maintained, which provided the possibility for its clinical application in the future.
【作者單位】： 中國醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所北京協(xié)和醫(yī)學(xué)院基礎(chǔ)學(xué)院病理系;
【分類號(hào)】：R392

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 汪華僑,范海虹,徐杰,李光武,袁群芳,謝瑤,姚志彬;抗氧化劑TA99系列治療阿爾茨海默病的實(shí)驗(yàn)依據(jù)(英文)[J];中國臨床康復(fù);2005年13期

2 錢云;張志s，

本文編號(hào)：1850371