Cell-surface antigens of melanoma recognized by human monoclonal antibodies.
Cell-surface antigens of melanoma recognized by human monoclonal antibodies.
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人类单克隆抗体识别的黑色素瘤细胞表面抗原。
DOI：
10.1073/pnas.84.8.2416
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发表时间：
1987
影响因子：
11.1
通讯作者：
Old,LJ
中科院分区：
文献类型：
--
作者：
Yamaguchi,H;Furukawa,K;Fortunato,SR;Livingston,PO;Lloyd,KO;Oettgen,HF;Old,LJ
人单克隆抗体（mAb）来源于黑素瘤患者的淋巴结淋巴细胞和外周血淋巴细胞（PBL）。比较了产生人mAb的四种方法：与人[LICR-LON-HMy-2（LICR-2）]或小鼠（NS-1）细胞融合; EB病毒（EBV）转化;以及EBV转化后NS-1融合。NS-1与淋巴结淋巴细胞的融合导致了比LICR-2融合更高数量的生长杂交体。NS-1或LICR-2与PBL的融合几乎没有获得杂种。EBV转化淋巴结和外周血淋巴细胞具有相同的效率，和抗体筛选的增殖培养物的产量超过10- 30倍以上，通过融合技术获得的。然而，一旦抗体产生的文化已经确定，EBV转化细胞的稳定性和克隆性差于NS-1杂交细胞。为了联合收割机结合两种方法的优点，将EBV转化细胞的培养物与NS-1融合;分离出显示出旺盛生长、可克隆性和稳定抗体分泌的杂交克隆。对这些克隆中的6个产生的mAb的详细特异性分析表明检测到1类（独特）黑素瘤抗原、3类黑素瘤抗原和4种神经节苷脂抗原（GD 3、GM 3和另外两种尚未表征的嗜异性抗原）。
Human monoclonal antibodies (mAbs) were derived from lymph node lymphocytes and peripheral blood lymphocytes (PBL) from patients with melanoma. Four methods for generating human mAbs were compared: fusion with human [LICR-LON-HMy-2 (LICR-2)] or mouse (NS-1) cells; transformation by Epstein-Barr virus (EBV); and EBV transformation followed by NS-1 fusion. NS-1 fusion with lymph node lymphocytes resulted in a higher number of growing hybrids than LICR-2 fusion. Virtually no hybrids were obtained from NS-1 or LICR-2 fusions with PBL. EBV transformed lymphocytes from lymph node and peripheral blood with equal efficiency, and the yield of proliferating cultures for antibody screening was more than 10- to 30-fold greater than that obtained by fusion techniques. However, once antibody-producing cultures had been identified, stability and clonability of EBV-transformed cells were poorer than that of NS-1 hybrid cells. To combine the strengths of both methods, cultures of EBV-transformed cells were fused with NS-1; and hybrid clones were isolated that showed vigorous growth, clonability, and stable antibody secretion. Detailed specificity analysis of the mAbs produced by six of these clones indicated detection of a class 1 (unique) melanoma antigen, a class 3 melanoma antigen, and four ganglioside antigens (GD3, GM3, and two other, as yet uncharacterized, heterophile antigens).