Detection and Quantitation of Human Ia‐Type Antigens with Iodinated Protein A and Specific Purification of Antibodies Against Ia‐Type Alloantigens
Detection and Quantitation of Human Ia‐Type Antigens with Iodinated Protein A and Specific Purification of Antibodies Against Ia‐Type Alloantigens
复制标题
用碘化蛋白 A 检测和定量人 Ia 型抗原以及抗 Ia 型同种抗原抗体的特异性纯化
DOI：
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发表时间：
1977
影响因子：
3.7
通讯作者：
H. Wigzell
中科院分区：
文献类型：
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作者：
D. Baron;P. Wernet;F. Schunter;H. Wigzell
在新鲜提取的人外周血白细胞、培养的人细胞系或用戊二醛或甲醛固定的淋巴细胞表面，使用碘化蛋白A与特定抗体-Ia-抗原复合体结合，可以很容易地检测和定量针对特定人类Ia型抗原的抗体。同样的原理也可以用于检测人类淋巴细胞上的Ia同种异体抗原，当用已知含有针对移植抗原的抗体的特定抗血清进行检测时。这些抗Ia-同种异体抗原抗体是通过在pH 6.3的DEAE-纤维素上离子交换层析和甲醛固定的Ia-同种异体抗原纯合子细胞株上的特异性吸附，然后在pH 3.0的等渗柠檬酸缓冲液中洗脱的两步法得到纯化的。通过这种方法，可以实现大约90倍的提纯。经过这样的纯化后，高度浓缩的抗体部分仍然选择性地与Ia抗原系统的一种特异性反应。
Antibodies directed against specific human Ia‐type antigens can easily be detected and quantitated by an improved radioimmunoassay using iodinated protein A bound to a specific antibody‐Ia‐antigen complex on the surface of freshly drawn peripheral human leukocytes, cultured human cell lines, or lymphoid cells fixed with glutardialdehyde or formaldehyde. The same principle can also be used for the detection of Ia alloantigens on human lymphocytes when testing them with specific antisera known to contain antibodies against transplantation antigens. These anti‐Ia‐alloantigen antibodies had been purified by a two‐step procedure involving ion‐exchange chromatography on DEAE‐cellulose at pH 6.3 and the specific absorption on formaldehyde‐fixed Ia‐alloantigen‐carrying homozygous cell lines, followed by elution of these antibodies with isotonic citrate buffer at pH 3.0. In this way an about 90‐fold purification could be achieved. After such a purification the highly enriched antibody fraction still reacted selectively with one specificity of the Ia antigen system.