PURIFICATION WITH MONOCLONAL ANTIBODY OF A PREDOMINANT LEUKOCYTE COMMON ANTIGEN AND GLYCOPROTEIN FROM RAT THYMOCYTES
PURIFICATION WITH MONOCLONAL ANTIBODY OF A PREDOMINANT LEUKOCYTE COMMON ANTIGEN AND GLYCOPROTEIN FROM RAT THYMOCYTES
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DOI：
10.1002/eji.1830090212
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发表时间：
1979-01-01
影响因子：
5.4
通讯作者：
WILLIAMS, AF
中科院分区：
文献类型：
--
作者：
SUNDERLAND, CA;MCMASTER, WR;WILLIAMS, AF
从大鼠胸腺细胞中纯化出一种白细胞共同（L - C）抗原，该抗原在兔抗大鼠胸导管淋巴细胞血清中作为免疫原可能具有优势。首先，通过扁豆凝集素亲和层析和在脱氧胆酸盐中的凝胶过滤，以400 - 900倍的纯化倍数制备出表观分子量为100,000的抗原片段。然后用该组分免疫小鼠，并通过细胞融合制备出分泌抗L - C抗原抗体的杂交骨髓瘤细胞系。该抗体用于亲和层析，与胸腺细胞相比，纯化倍数达到1400倍，从而得到纯的L - C抗原。L - C抗原是大鼠胸腺细胞的一种主要膜糖蛋白，通过在十二烷基硫酸钠 - 聚丙烯酰胺凝胶上电泳测定，其表观分子量为150,000。该抗原是用高碘酸 - 希夫试剂对糖类染色强烈的3种胸腺细胞糖蛋白中的1种。它存在于超过95%的胸腺细胞、骨髓细胞和胸导管淋巴细胞上。
A leukocyte-common (L-C) antigen which can be dominant as an immunogen in rabbit anti-rat thoracic duct lymphocyte serum was purified from rat thymocytes. Initially, an antigenic fragment of 100,000 apparent MW was prepared at 400- to 900-fold purification by lentil lectin affinity chromatography and gel filtration in deoxycholate. Mice were then immunized with this fraction, and a hybrid myeloma cell line secreting antibody to the L-C antigen was prepared by cell fusion. This antibody was used for affinity chromatography and gave pure L-C antigen at 1400-fold purification compared with thymocytes. The L-C antigen is a major membrane glycoprotein of rat thymocytes and has an apparent MW of 150,000 as determined by electrophoresis on polyacrylamide gels in sodium dodecyl sulfate. The antigen constitutes 1 of the 3 thymocyte glycoproteins which stain intensely for carbohydrate with periodic acid Schiff stain. It is present on > 95% of thymocytes, bone marrow cells and thoracic duct lymphocytes.