Clausen H, Levery S B, Nudelman E, Tsuchiya S, Hakomori S
Proc Natl Acad Sci U S A. 1985 Feb;82(4):1199-203. doi: 10.1073/pnas.82.4.1199.
The IgG2a monoclonal antibody TH-1, which reacts specifically with blood group A1 but with neither A2 nor O erythrocytes, has been established. The antibody reacted only with A1 erythrocytes in hemagglutination and antibody absorption assays; it did not react with A2 erythrocytes, even after trypsin or sialidase treatment. This antibody detected, on TLC immunostaining, a series of glycolipids from A1 erythrocytes but virtually none or very weak bands from A2 erythrocytes. It did not react with type 1 or type 2 chain A, or with globo-A. The simplest reactive component was isolated from a previously assigned Ab fraction by HPTLC of acetylated compounds. The structure of the reactive component was characterized by 1H NMR spectroscopy, methylation analysis, and enzymatic degradation, as shown below: (Formula: see text). The structure is essentially a repetitive A epitope attached to type 2 chain and is hereby called type 3 chain A. The determinant can be carried on extended and/or branched structures, but it was not detectable in glycoproteins. The structure was characteristic of A1 erythrocytes and present in only trace amounts in A2 erythrocytes. The precursor H (Fuc alpha 1----2Gal beta 1----3GalNAc alpha 1----3[Fuc alpha 1----2]Gal beta 1----4GlcNAc beta 1----R; type 3 chain H) was present in greater quantity in A2 erythrocytes than in A1 erythrocytes, but it was absent in both O and B erythrocytes. The A1 transferase apparently can transfer alpha-GalNAc to type 3 chain H, while the A2 transferase may not have this ability.
已制备出IgG2a单克隆抗体TH - 1,它能与A1血型红细胞特异性反应,但不与A2或O型红细胞反应。在血凝和抗体吸收试验中,该抗体仅与A1红细胞反应;即使经胰蛋白酶或唾液酸酶处理后,它也不与A2红细胞反应。通过薄层层析免疫染色,此抗体能检测到来自A1红细胞的一系列糖脂,但来自A2红细胞的条带几乎没有或非常微弱。它不与1型或2型A链或球型A反应。通过对乙酰化化合物进行高效薄层层析,从先前指定的抗体组分中分离出了最简单的反应性成分。如下所示,通过1H核磁共振光谱、甲基化分析和酶促降解对反应性成分的结构进行了表征:(分子式:见正文)。该结构本质上是一个连接在2型链上的重复A表位,在此称为3型链A。该决定簇可存在于延伸和/或分支结构上,但在糖蛋白中无法检测到。该结构是A1红细胞的特征性结构,在A2红细胞中仅微量存在。前体H(Fucα1----2Galβ1----3GalNAcα1----3[Fucα1----2]Galβ1----4GlcNAcβ1----R;3型链H)在A2红细胞中的含量高于A1红细胞,但在O型和B型红细胞中均不存在。A1转移酶显然可以将α - GalNAc转移到3型链H上,而A2转移酶可能没有这种能力。