Kopitar-Jerala N, Bestagno M, Fan X, Novak-Despot D, Burrone O, Kos J, Skrk J, Gubensek F
Department of Biochemistry and Molecular Biology, Jozef Stefan Institute, Ljubljana, Slovenia.
Pflugers Arch. 2000;439(3 Suppl):R79-80.
A chimeric mouse-human antibody has been created that recognizes an antigen found on breast cancer cells and melanoma cells. Immunoglobulin constant domains of mouse monoclonal antibody CDI 315B Cgamma1 and CK, were substituted by the human Cgamma1 and Ckappa. The CDI 315B variable heavy and light chain regions were PCR amplified from hybridoma RNA and sequenced. Mouse variable VH and VL regions were joint to human IgG1 and kappa constant regions and subcloned into pcDNA3 expression vectors. The Sp2/0 murine myeloma cells were transfected with expression vectors pcDNA3L and pcDNA3H and the reactivity of chimeric antibodies was tested by indirect ELISA using B16F1 murine melanoma cells as well as MCF7 human breast cancer cells, as antigen.
已构建出一种嵌合型小鼠-人抗体，它能识别在乳腺癌细胞和黑色素瘤细胞上发现的一种抗原。小鼠单克隆抗体CDI 315B的免疫球蛋白恒定结构域Cgamma1和CK被人Cgamma1和Ckappa取代。从杂交瘤RNA中通过PCR扩增CDI 315B可变重链和轻链区域并进行测序。将小鼠可变VH和VL区域与人IgG1和kappa恒定区域连接，并亚克隆到pcDNA3表达载体中。用表达载体pcDNA3L和pcDNA3H转染Sp2/0鼠骨髓瘤细胞，并以B16F1鼠黑色素瘤细胞以及MCF7人乳腺癌细胞作为抗原，通过间接ELISA检测嵌合抗体的反应性。
