Charrin Stéphanie, Palmulli Roberta, Billard Martine, Clay Denis, Boucheix Claude, Van Niel Guillaume, Rubinstein Eric
Centre d'Immunologie et des Maladies Infectieuses, Inserm, CNRS, Sorbonne Université, CIMI-Paris, 75013 Paris, France.
Centre National de la Recherche Scientifique, Structure and Membrane Compartments, Institut Curie, Paris Sciences & Lettres Research University, UMR144, 75005 Paris, France.
Antibodies (Basel). 2020 Jul 2;9(3):29. doi: 10.3390/antib9030029.
CD63, a member of the tetraspanin superfamily, is used as a marker of late endosomes and lysosome-related organelles, as well as a marker of exosomes. Here, we selected rare isotype variants of TS63 by sorting hybridoma cells on the basis of their high expression of surface immunoglobulins of the IgG2a and IgG2b subclass. Pure populations of cells secreting IgG2a and IgG2b variants of TS63 (referred to as TS63a and TS63b) were obtained using two rounds of cell sorting and one limited dilution cloning step. We validate that these new TS63 variants are suitable for co-labeling with mAb of the IgG1 subclass directed to other molecules, using anti mouse subclass antibodies, and for the labeling of exosomes through direct binding to protein A-coated gold particles. These mAbs will be useful to study the intracellular localization of various proteins and facilitate electron microscopy analysis of CD63 localization.
CD63是四跨膜蛋白超家族的成员，用作晚期内体和溶酶体相关细胞器的标志物，以及外泌体的标志物。在此，我们通过基于IgG2a和IgG2b亚类表面免疫球蛋白的高表达对杂交瘤细胞进行分选，筛选出了TS63的罕见同种型变体。通过两轮细胞分选和一轮有限稀释克隆步骤，获得了分泌TS63的IgG2a和IgG2b变体（称为TS63a和TS63b）的纯细胞群体。我们验证了这些新的TS63变体适用于与针对其他分子的IgG亚类单克隆抗体共同标记，使用抗小鼠亚类抗体，以及通过直接结合蛋白A包被的金颗粒对外泌体进行标记。这些单克隆抗体将有助于研究各种蛋白质的细胞内定位，并促进对CD63定位的电子显微镜分析。
