SEREX方法筛选人胎盘抗原

SEREX Screening of Human Placenta Antigens

目的筛选并获得来自人胎盘有潜在应用价值的抗原。方法基于血清学筛选重组cDNA表达文库(serological analysis of recombinant cDNA expression library,SEREX)技术,制备人绒毛组织免疫的兔血清,利用该免疫血清筛选人胎盘组织cDNA表达文库,通过两轮筛选得到免疫反应阳性的单克隆重组噬菌体并测序,并对所得到的序列进行生物信息学分析。结果经过两轮筛选共得到69个阳性反应克隆,序列分析发现它们代表12种不同的基因序列,其中功能已知基因9个,功能未知基因3个,进一步分析发现绒毛膜生长催乳激素基因(CSH1和CSH2)有57个阳性反应克隆,占总数的82.6%,可能是一个具有重要功能的抗原基因,所筛选到的其他基因与胚胎生长发育相关。结论SEREX技术筛选胎盘抗原是行之有效的,本实验筛选出的抗原基因对研究胚胎生长发育具有一定的意义。

Objective To screen and obtain the potential human placenta antigens for the further application with serological analysis of recombinant cDNA expression library （SEREX）. Methods SEREX technique with some modifications was applied. In brief, immune sera from rabbit immunized with human chorionic tissue were used to screen human placenta tissue cDNA expression library. Positive clone plaques were obtained after two rounds of screen. And the positive clone plaques were purified and sequenced. BLAST software was applied to comparison the obtained sequences with those found in GenBank for bioinformatic analysis. Results 69 positive clones were obtained from two rounds of screen. They were derived from 12 different genes, 9 of these were genes of known biology function, and 3 were genes of unknown biology function depending on the sequence analysis. Among the positive clones, chorionic somatomammotropin hormone 1 and 2 genes （CSH1 and CSH2） were found in 57 of positive clones （82. 6%）. This implied that the CSH1 and CSH2 might be the gene of encoding the important antigen and other genes obtained were related to the development of embryo. Conclusion Modified xenogeneic immune SEREX technology is a very effective method to screen and isolate human placenta antigens. These antigens identified from this study might contribute to clarify the process of embryo development.