Loirat M J, Dahr W, Muller J Y, Blanchard D
Centre Régional de Transfusion Sanguine, Nantes, France.
Transfus Med. 1994 Jun;4(2):147-55. doi: 10.1111/j.1365-3148.1994.tb00256.x.
Six new murine monoclonal antibodies (mAbs) directed to the erythrocyte membrane glycophorins C (GPC) and D (GPD) were obtained from splenocytes of different BALB/c mice immunized with human red blood cells, and fully characterized. The mAbs were selected by agglutination tests with control and Gerbich-negative cells, and by immunoblotting analysis. They showed specificity for the N-terminal domain(s) of GPC (and GPD) and were classified into three categories by competitive analysis using 125I-labelled antibodies and real-time biospecific interaction. The first group (NaM10-7G11, NaM70-1G4 and NaM77-7B6) compete for epitope(s) located at the N-terminal portion of GPC. Agglutination-inhibition tests revealed that the 7G11 epitope involves the amino group of Met1 and sialic acid residue(s) whereas the 1G4 and 7B6 epitopes contain O-glycans. NaM89-2G11 belongs to a second group; its epitope is located in a region including Glu17, Asp19 and (an) O-glycan(s). The third group comprises mAbs NaM19-3C4 and NaM98-3C1 which bind to both GPC and GPD in proximity of the binding site of human anti-Ge:3 antibodies. In addition, mAb 3C4 (anti-GPC/GPD) was found to bind to approximately 125,000 sites per red cell. Considering that the ratio of the GPC to GPD is about 3-4 to 1, the number of GPC and GPD molecules was estimated as 95,000 and 35,000, respectively.
从用人类红细胞免疫的不同BALB/c小鼠的脾细胞中获得了六种针对红细胞膜糖蛋白C（GPC）和D（GPD）的新型鼠单克隆抗体（mAb），并对其进行了全面表征。通过与对照细胞和Gerbich阴性细胞的凝集试验以及免疫印迹分析来选择这些单克隆抗体。它们对GPC（和GPD）的N端结构域具有特异性，并通过使用125I标记抗体的竞争分析和实时生物特异性相互作用分为三类。第一组（NaM10-7G11、NaM70-1G4和NaM77-7B6）竞争位于GPC N端部分的表位。凝集抑制试验表明，7G11表位涉及Met1的氨基和唾液酸残基，而1G4和7B6表位含有O-聚糖。NaM8۹-2G11属于第二组；其表位位于包括Glu17、Asp19和（一个）O-聚糖的区域。第三组包括mAb NaM19-3C4和NaM98-3C1，它们在人抗Ge:3抗体的结合位点附近与GPC和GPD都结合。此外，发现mAb 3C4（抗GPC/GPD）与每个红细胞上约125,000个位点结合。考虑到GPC与GPD的比例约为3 - 4比1，估计GPC和GPD分子的数量分别为95,000和35,000。
