多发性骨髓瘤MMSA-1抗原的二级结构预测及多克隆抗体的制备  

Predicting the secondary structure and preparing polyclonal antibody for a novel antigen MMSA-1 in multiple myeloma

作　　者：周芙玲[1] 张王刚[1] 王佰言[1] 张鹏宇[1] 古流芳[1] 张文娟[1] 蒙昕[1] 田纬[1] 

机构地区：[1]西安交通大学医学院第二附属医院血液内科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2009年第4期449-452,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No .30700337)~~

摘　　要：目的运用生物信息学技术研究人多发性骨髓瘤MMSA-1(GenBank:AY952881)基因表达蛋白的二级结构和B细胞抗原表位,并制备多克隆抗体。方法应用DNAStar Protean软件提供的模块分析和预测MMSA-1蛋白的二级结构和B细胞抗原表位。采用多通道同步固相全自动合成系统合成15肽,免疫健康新西兰成年家兔,制备相应的多克隆抗体。结果MMSA-1蛋白含有较多的β折叠和转角结构。综合柔韧性、亲水性、抗原性、表面可能性、二级结构、偶联难易及实验难度等因素,选取了MMSA-1蛋白序列318～333位作为抗原表位,即SSSLLPQSPAPTEHL,15肽合成产物经HPLC纯化和质谱检测,纯度达到98.65%,序列正确。免疫兔子获得血清抗体采用层析柱纯化,ELISA检测效价:1∶6000。结论本研究为基于人MMSA-1抗原表位手段的免疫治疗研究提供了理论依据。Objective Prediction was made for a novel antigen （multiple myeloma special antigen） MMSA-1 in human multiple myeloma by using bioinformatics. We predicted the secondary structure and B cell epitopes, and elicited specific antibody using peptide-microspheres and adjuvants. Methods The secondary structure and B cell epitopes of MMSA-1 protein were determined by graphic analysis using the Protean module （DNAstar Inc.）. Antigenic peptides were synthesized as the antigen with Fmoc/PyBOP method. Adult New Zealand rabbits were immunized by injecting the chemical coupling of synthetic peptides-KLH to obtain antibody. Results MMSA-1 protein contained more β sheet and turn regions. Antigenic peptides （SSSLLPQSPAPTEHL） were purified above 98.65% by high performance liquid chromatography and validated by mass spectrometric detection. The immunized sera analyzed with ELISA were 1 ： 6 000 after purification using a peptide affinity column. Conclusion The prediction of tumor antigens provides thereotical evidence for the immunotherapy of multiple myeloma. Moreover, antibody preparation is useful for further study.

关 键 词：多发性骨髓瘤 MMSA-1 二级结构 抗原表位 多克隆抗体 

分 类 号：R730.2[医药卫生—肿瘤]