鼠抗人Syndecan-1分子功能性单克隆抗体的制备及其生物学特性

Preparation of functional anti-human Syndecan-1 monoclonal antibody and its biological characteristics

制备鼠抗人Syndecan-1(CD138)分子的功能性单克隆抗体,研究其对高表达CD138分子细胞的生物学效应。以人多发性骨髓瘤(MM)细胞株8226作为免疫原,8226和B淋巴瘤细胞株Daudi作为检测细胞株,利用B淋巴细胞杂交瘤技术制备单克隆抗体。以快速定性试纸分析法鉴定单抗所属的IgG亚类,采用间接免疫荧光标记法及竞争抑制实验分析单抗的亲和力和特异性,以高表达Syndecan-1的人多发性骨髓瘤细胞株XG-1为靶细胞,分析单抗对其生长的影响。结果:成功地获得1株鼠抗人Syndecan-1分子的功能性单克隆抗体杂交瘤细胞株(克隆4B3),经体外长期传代培养,液氮冻存半年以上反复复苏良好,分泌特异性抗体性能稳定。4B3分泌的单克隆抗体识别位点与单抗BB4识别位点相同或相近,重链为IgG1亚类,轻链为κ型,纯化后腹水中单克隆抗体蛋白的得率为2.9 mg/ml,加入4B3单克隆抗体后的XG-1细胞体外生长受到抑制,并可用于神经胶质瘤细胞的免疫组织化学检测。由此表明,4B3为功能性抗人Syndecan-1单克隆抗体,具有一定的基础研究和临床应用前景,这对进一步研究Syndecan-1分子的生物学特性也具有重要的价值。

To obtain mouse anti-human Syndencan-1 monoclonal antibody（mAb） cell fusion, and screening, immunofluorescence analysis, immunoblotting and competition test were to he used. By indirect immunofluorescence assay the expression of Syndecan-1 on 8226, U266, XG-1 , XG-2, Daudi and Jurkat cell lines were investigated. Cell counting and trypan blue staining were used to analyze the inhibitory effects of mAb on the proliferation of XG-1 cell. The mouse anti-human Syndecan-1 hybridoma cell line 4B3 was thus obtained. The subclass of mAb produced by cell line 4B3 was mouse IgG1 and the 4B3 mAb could recognize the identical or similar epitope with PE-conjugated positive control mAb. This mAb could inhibit the growth of XG-1 cell. It is concluded that 4B3 mAb is a specific and functional anti-Syndeean-1 mAb and has high affinity for its ligand. It may be of significant value for basic studies and for clinical applications.