篩選及鑒定IgG型單克隆抗-D噬菌體克隆的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-08-21 08:07
【摘要】：目的利用隨機(jī)噬菌體12肽庫(kù)篩選出血型D抗原的模擬多肽并驗(yàn)證。方法采用Ig G型單克隆抗-D篩選噬菌體隨機(jī)12肽庫(kù),測(cè)定每輪篩選后洗脫物與抗-Rh D的結(jié)合情況。對(duì)經(jīng)特異性篩選得到的陽(yáng)性噬菌體克隆進(jìn)行DNA序列測(cè)定,將推導(dǎo)出的12肽氨基酸序列進(jìn)行同源性比較,通過(guò)凝集抑制試驗(yàn)驗(yàn)證陽(yáng)性噬菌體與Ig G型單克隆抗-D的反應(yīng)。結(jié)果隨著篩選輪次增加,噬菌體洗脫物與抗-Rh D的結(jié)合力逐漸增強(qiáng)。經(jīng)過(guò)4輪淘洗后,得到8個(gè)親和力較強(qiáng)且具有較高重合率的保守區(qū)域的12肽序列。凝集抑制實(shí)驗(yàn)表明具有相同氨基酸序列的陽(yáng)性噬菌體對(duì)Ig G型單克隆抗-D與Rh D陽(yáng)性紅細(xì)胞的凝集反應(yīng)具有抑制作用。結(jié)論隨機(jī)噬菌體12肽庫(kù)篩選得到的血型D抗原模擬多肽,為Rh D結(jié)構(gòu)與功能研究及研制針對(duì)Rh D新生兒溶血病的新型診斷抗原、制備特異疫苗以及探索新的治療方法提供實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Objective to screen and verify the mimic peptide of blood group D antigen using random phage 12 peptide library. Methods the phage random 12 peptide library was screened by Ig G monoclonal anti-D and the binding of the eluate to anti-Rh D was determined after each screening. The DNA sequence of positive phage clones was determined by specific screening. The deduced 12-peptide amino acid sequences were compared with each other. The agglutination inhibition test was used to verify the reaction between the positive phage and IgG monoclonal anti-D. Results with the increase of screening times, the binding ability of phage elution and anti-Rh D increased gradually. After four rounds of washing, the 12 peptide sequences of 8 conserved regions with strong affinity and high coincidence rate were obtained. The agglutination inhibition experiment showed that the positive phage with the same amino acid sequence could inhibit the agglutination of IgG monoclonal anti-D and Rh D positive erythrocytes. Conclusion the mimic peptide of blood group D antigen obtained from random phage 12 peptide library is a novel diagnostic antigen for neonatal hemolytic disease of Rh D, which is used to study the structure and function of Rh D and to develop a novel antigen for diagnosis of Rh D hemolytic disease. Preparation of specific vaccines and exploration of new therapeutic methods provide experimental basis.
【作者單位】： 深圳市血液中心;
【基金】：深圳市衛(wèi)生計(jì)生系統(tǒng)科研項(xiàng)目(201502008)
【分類號(hào)】：R446.6
，
本文編號(hào)：2195082
[Abstract]:Objective to screen and verify the mimic peptide of blood group D antigen using random phage 12 peptide library. Methods the phage random 12 peptide library was screened by Ig G monoclonal anti-D and the binding of the eluate to anti-Rh D was determined after each screening. The DNA sequence of positive phage clones was determined by specific screening. The deduced 12-peptide amino acid sequences were compared with each other. The agglutination inhibition test was used to verify the reaction between the positive phage and IgG monoclonal anti-D. Results with the increase of screening times, the binding ability of phage elution and anti-Rh D increased gradually. After four rounds of washing, the 12 peptide sequences of 8 conserved regions with strong affinity and high coincidence rate were obtained. The agglutination inhibition experiment showed that the positive phage with the same amino acid sequence could inhibit the agglutination of IgG monoclonal anti-D and Rh D positive erythrocytes. Conclusion the mimic peptide of blood group D antigen obtained from random phage 12 peptide library is a novel diagnostic antigen for neonatal hemolytic disease of Rh D, which is used to study the structure and function of Rh D and to develop a novel antigen for diagnosis of Rh D hemolytic disease. Preparation of specific vaccines and exploration of new therapeutic methods provide experimental basis.
【作者單位】： 深圳市血液中心;
【基金】：深圳市衛(wèi)生計(jì)生系統(tǒng)科研項(xiàng)目(201502008)
【分類號(hào)】：R446.6
，
本文編號(hào)：2195082
本文鏈接：http://sikaile.net/xiyixuelunwen/2195082.html
最近更新
教材專著
