Kreutz F T, Xu D Z, Suresh M R
Faculty of Pharmacy and Pharmaceutical Science, University of Alberta, Edmonton, Canada.
Hybridoma. 1998 Jun;17(3):267-73. doi: 10.1089/hyb.1998.17.267.
Bispecific monoclonal antibodies (bsMAbs) are unique molecules incorporating two different paratopes in a single antibody molecule. BsMAbs can be useful in different areas of research as well in clinical applications. Traditionally, bsMAbs are produced by hybrid-hybridomas that are generated by the fusion of two pre-established hybridomas. The development of such hybrid-hybridomas can be difficult and time-consuming. Here, we introduce a new technique to generate such hybrids, electro-FACS-fusion. In this procedure, before the electrofusion, one of the hybridomas is labeled with fluorescein isothiocyanate (FITC) and the other with tetramethylrhodamine isothiocyanate (TRITC). The mixture of cells is then electrofused, and cells exhibiting dual fluorescence are selected by fluorescence activated cell sorting (FACS). The fused cells are directly plated in microplates for clonal growth. Using this technique, we produced three new hybrid-hybridomas secreting bsMAb that could be used for the next generation of immunoassays.
双特异性单克隆抗体（bsMAb）是一类独特的分子，在单个抗体分子中包含两种不同的抗原结合位点。bsMAb在不同的研究领域以及临床应用中都可能有用。传统上，bsMAb是由杂交杂交瘤产生的，杂交杂交瘤是通过将两种预先建立的杂交瘤融合而产生的。这种杂交杂交瘤的开发可能困难且耗时。在此，我们介绍一种产生此类杂交体的新技术，即电FACS融合。在该过程中，在电融合之前，其中一种杂交瘤用异硫氰酸荧光素（FITC）标记，另一种用异硫氰酸四甲基罗丹明（TRITC）标记。然后将细胞混合物进行电融合，并通过荧光激活细胞分选（FACS）选择显示双重荧光的细胞。将融合细胞直接接种到微孔板中进行克隆生长。使用该技术，我们产生了三种分泌bsMAb的新杂交杂交瘤，可用于下一代免疫测定。
