【摘要】：目的利用隨機噬菌體12肽庫篩選出血型D抗原的模擬多肽并驗證。方法采用Ig G型單克隆抗-D篩選噬菌體隨機12肽庫,測定每輪篩選后洗脫物與抗-Rh D的結合情況。對經(jīng)特異性篩選得到的陽性噬菌體克隆進行DNA序列測定,將推導出的12肽氨基酸序列進行同源性比較,通過凝集抑制試驗驗證陽性噬菌體與Ig G型單克隆抗-D的反應。結果隨著篩選輪次增加,噬菌體洗脫物與抗-Rh D的結合力逐漸增強。經(jīng)過4輪淘洗后,得到8個親和力較強且具有較高重合率的保守區(qū)域的12肽序列。凝集抑制實驗表明具有相同氨基酸序列的陽性噬菌體對Ig G型單克隆抗-D與Rh D陽性紅細胞的凝集反應具有抑制作用。結論隨機噬菌體12肽庫篩選得到的血型D抗原模擬多肽,為Rh D結構與功能研究及研制針對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驗基礎。
[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)生計生系統(tǒng)科研項目(201502008)
【分類號】：R446.6
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本文編號：2195082