Du Xue-mei, Tang Li, Liu Xiao-lan, Chen Yong, Gao Jian-en, He Fu-chu, Sun Qi-hong
Department of Immunology, Beijing Institute of Radiation Medicine, Beijing 100850, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jul;22(4):517-20.
To prepare and characterize monoclonal antibody (mAb) against human LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin) protein.
BALB/c mice were immunized with prokaryotically expressed human LSECtin protein. The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. Reaction of mAb to LSECtin antigen was characterized by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM.
Eight hybridoma cells secreting mAbs were established. The isotypes of the mAbs were IgG. Ascites titers were between 1:10(6) - 1:10(7). All the mAbs recognized human LSECtin protein on LSECtin-transfected 3T3 cells and six of the mAbs specifically recognized liver sinusoidal endothelial cells.
Eight anti-LSECtin mAbs have been obtained. The characterization of the mAbs indicate that they show fine specificity by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM, which can provide a powerful reagent for the functional study of LSECtin.
制备并鉴定抗人LSECtin（肝脏和淋巴结窦状内皮细胞C型凝集素）蛋白的单克隆抗体（mAb）。
用原核表达的人LSECtin蛋白免疫BALB/c小鼠。将免疫小鼠的脾细胞与鼠骨髓瘤细胞（Sp2/0）融合，然后通过间接ELISA筛选mAb阳性杂交瘤细胞。通过蛋白质印迹法、间接免疫荧光染色、免疫组织化学染色和流式细胞术对mAb与LSECtin抗原的反应进行鉴定。
建立了8株分泌mAb的杂交瘤细胞。mAb的亚型为IgG。腹水效价在1:10(6) - 1:10(7)之间。所有mAb均能识别LSECtin转染的3T3细胞上的人LSECtin蛋白，其中6株mAb能特异性识别肝窦状内皮细胞。
已获得8株抗LSECtin mAb。mAb的鉴定表明，它们通过蛋白质印迹法、间接免疫荧光染色、免疫组织化学染色和流式细胞术表现出良好的特异性，可为LSECtin的功能研究提供有力试剂。
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