Sagisaka K, Iwasa M, Umezu C
Tohoku J Exp Med. 1977 Jul;122(3):267-74. doi: 10.1620/tjem.122.267.
Anti-M and -N antibodies are usually prepared from rabbit immunized with human O.M or O.N red cells. The crude antisera must be adsorbed with non-corresponding red cells to remove heteroagglutinin. To specify the crude antisera, we have employed dextran gel (DEAE-Sephadex A-50) as an immunoadsorbent, which was mixed with M or N group substance prepared by the phenol-saline method, and was fixed with formaldehyde. The substance-coupled gel was added to the corresponding crude antiserum and washed with cold saline. Then, the gel added with an equal volume of saline was heated at 55 degrees C for 10 min and applied on a funnel with slight evacuating. By this method, specific anti-M and -N antibodies (titer 1:16 to 1:32) were prepared, but the antibody was not eluted from the gel reacted with IgM fraction. The recovery rate and specificity of the eluted antibodies are discussed.
抗-M和抗-N抗体通常由用人O.M或O.N红细胞免疫的兔子制备。粗抗血清必须用不相应的红细胞吸附以去除异凝集素。为了鉴定粗抗血清,我们使用葡聚糖凝胶(DEAE-葡聚糖A-50)作为免疫吸附剂,将其与用酚盐水法制备的M或N组物质混合,并用甲醛固定。将物质偶联凝胶加入相应的粗抗血清中,并用冷盐水洗涤。然后,将加入等体积盐水的凝胶在55℃加热10分钟,并应用于轻微抽气的漏斗上。通过这种方法,制备了特异性抗-M和抗-N抗体(效价为1:16至1:32),但抗体未从与IgM组分反应的凝胶中洗脱出来。讨论了洗脱抗体的回收率和特异性。