Baldwin C I, Denham D A
Dept. of Medical Parasitology, LSHTM, U.K.
Immunology. 1994 Jan;81(1):155-60.
Although the cat is an important model for a number of human diseases little is known of its basic humoral immunity. In this study we have isolated three subpopulations of IgG from cat serum by DEAE anion-exchange chromatography and protein A affinity chromatography. Subpopulation 3 did not bind to DEAE equilibrated in 2 mM phosphate buffer pH 8.0 while subpopulations 1 and 2 were eluted, as a mixed population, with a salt gradient between 3 mM and 25 mM. When this fraction was loaded onto protein A-Sepharose two distinct peaks were always generated in a pH gradient (pH 8.0-2.1). The first of these, subpopulation 2, was eluted on average at pH 4.3 while the second, subpopulation 1, was eluted on average at pH 3.4. Subpopulations 1 and 2 had similar immunoelectrophoretic mobilities and isoelectric points while subpopulation 3 was more cathodic in both tests. Antisera produced in both mice and rabbits were rendered specific, by absorption of cross-reacting antibodies, for determinants on the heavy chain of each subpopulation as shown by Western blotting. The data suggest, but are not conclusive, that each of these subpopulations are distinct subclasses of IgG.
尽管猫是多种人类疾病的重要模型,但对其基本体液免疫的了解却很少。在本研究中,我们通过DEAE阴离子交换色谱法和蛋白A亲和色谱法从猫血清中分离出了IgG的三个亚群。亚群3在pH 8.0的2 mM磷酸盐缓冲液平衡的DEAE上不结合,而亚群1和2作为混合群体,在3 mM至25 mM的盐梯度下被洗脱。当将该级分加载到蛋白A-琼脂糖上时,在pH梯度(pH 8.0 - 2.1)中总是产生两个不同的峰。其中第一个,亚群2,平均在pH 4.3时被洗脱,而第二个,亚群1,平均在pH 3.4时被洗脱。亚群1和2具有相似的免疫电泳迁移率和等电点,而亚群3在两项测试中都更偏向阴极。如Western印迹所示,通过吸收交叉反应抗体,在小鼠和兔子中产生的抗血清对每个亚群重链上的决定簇具有特异性。数据表明,但尚无定论,这些亚群中的每一个都是IgG的不同亚类。