针对兔肺核糖核酸酶的单特异性抗体。

Monospecific antibodies to rabbit lung ribonucleases.

作者信息

Hastie A T

出版信息

J Biol Chem. 1981 Dec 10;256(23):12553-60.

Abstract

Ribonucleases were isolated from normal rabbit lung by chromatographic techniques. Two active fractions were separated by elution from a sulfopropyl-Sephadex column with 0.2 and 1.0 M NaCl. These were further purified over 2000-fold from the original crude homogenate. Both RNase preparations showed more than one protein possessing RNase activity in polyacrylamide gel electrophoresis with either sodium dodecyl sulfate or low pH buffer systems. The purified fraction eluted at 0.2 M NaCl was injected into a goat. The resulting antiserum inhibited the RNase activity of both RNase preparations. Immunoelectrophoresis revealed three noncross-reacting precipitin arcs with the 0.2 M NaCl-eluted RNase and two noncross-reacting precipitin arcs with the 1.0 M NaCl-eluted RNase. Highly significant RNase activity was recovered from the beta arc in the former and the delta arc in the latter. By immunoabsorbant chromatography, the antiserum was separated into three monospecific fractions, anti-alpha, anti-beta, and anti-delta. Anti-beta and anti-delta inhibited, respectively, 60 and 33% of the 0.2 M NaCl-eluted RNase activity. Anti-delta inhibited 93% of the 1.0 M NaCl-eluted RNase activity. RNase activity in rabbit serum was not inhibited by any of the monospecific antibody fractions.

摘要

采用色谱技术从正常兔肺中分离出核糖核酸酶。通过用0.2和1.0M氯化钠从磺丙基-葡聚糖凝胶柱上洗脱，分离出两个活性组分。它们从原始粗匀浆中进一步纯化了2000多倍。在十二烷基硫酸钠或低pH缓冲系统的聚丙烯酰胺凝胶电泳中，两种核糖核酸酶制剂均显示出不止一种具有核糖核酸酶活性的蛋白质。将在0.2M氯化钠处洗脱的纯化组分注射到山羊体内。所得抗血清抑制了两种核糖核酸酶制剂的核糖核酸酶活性。免疫电泳显示，0.2M氯化钠洗脱的核糖核酸酶有三条不交叉反应的沉淀弧，1.0M氯化钠洗脱的核糖核酸酶有两条不交叉反应的沉淀弧。在前一种情况中，从β弧中回收了高度显著的核糖核酸酶活性，在后一种情况中，从δ弧中回收了高度显著的核糖核酸酶活性。通过免疫吸附色谱法，抗血清被分离成三个单特异性组分，抗α、抗β和抗δ。抗β和抗δ分别抑制了0.2M氯化钠洗脱的核糖核酸酶活性的60%和33%。抗δ抑制了1.0M氯化钠洗脱的核糖核酸酶活性的93%。兔血清中的核糖核酸酶活性不受任何单特异性抗体组分的抑制。