| 摘要: |
| 将培养收获的MSB1细胞用磷酸盐缓冲液洗涤3次后,调整细胞浓度为2×106/mL,然后每毫升细胞悬液加2 IU木瓜蛋白酶和1 mmol/L的L-胱氨酸溶液0.1 mL,混匀后37 ℃作用1 h以分离MSB1细胞表面MAT-SA。对木瓜蛋白酶处理前后的MSB1细胞进行间接荧光抗体染色以检测MATSA的分离情况。获得的MATSA粗提物经超滤浓缩和Sephadex G-75凝胶纯化后,用SDS-PAGE检测纯化的结果。试验结果表明,从MSB1细胞表面分离到MATSA,经电泳获得了纯化的M ATSA,其相对分子质量约为35 ku。结果为MATSA单克隆抗体的制备及其相关的研究奠定了基础。 |
| 关键词: 马立克病肿瘤相关表面抗原 MATSA的分离与纯化 |
| DOI： |
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| 基金项目:国家自然科学基金项目（30371067）；霍英东教育基金项目（91033） |
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| Separation and identification and purification of Marek's disease tumor-associated surface antigen on MSB1 cells |
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| Abstract: |
| MSB1 cells were harvested and washed three times with phosphate buffer saline,and then the concentration was regulated to be 2×106/mL.2 IU papain and 1 mmol/L L-cystine 0.1 mL to 2×106 MSB1 cells were added and then incubated 1h in 37 ℃.Normal MSB1 and MSB1 treated with papin were detected with-indirect fluorescent antibody assay to indicate whether MATSA was removed from MSB1 cells.Crude extract of MATSA was concentrated and purified using ultrafiltration and gel filtration of Sephadex G-75.SDS-PAGE detected purification result detected which proved that MATSA had been removed from MBS1 cells by indirect fluorescent antibody assay.Detection of sodium dodecyl sulfate polyacrylamide gel electropheresis indicated that MATSA had been separated and purified.The molecular weight of purified MATSA was about 35 ku. |
| Key words: MSB1 marek’s disease tumor-associated surface antigen separati on and purification of MATSA |
