Adeyemi E O, Hodgson H J
Department of Medicine, Royal Postgraduate Medical School, London, UK.
J Immunol Methods. 1993 Sep 15;164(2):189-92. doi: 10.1016/0022-1759(93)90311-t.
In order to investigate the effect which the binding of anti-alpha 2-macroglobulin (anti-A2M) antibody has on subsequent antigen-antibody reaction between the complexed elastase and the solid phase anti-elastase antibody, elastase alpha 2-macroglobulin complex (EMC) was incubated with anti-A2M antibody and then extracted by a solid-phase bound rabbit anti-elastase antibody. A doubling dilution series of EMC generated dose related absorbance values. The critical factor permitting the immunological detection of A2M-bound elastase is the pre-incubation of anti-A2M antibody with EMC in solution. The assay exhibited a lower detection limit of 0.5 ng bound elastase per ml and EMC levels in serum samples from ten volunteers were significantly higher than in plasma (28 vs. 21 ng/ml, p < 0.05, Student's t test for paired samples). The EMC levels measured with this assay were essentially identical to those obtained when phenyl methyl sulfonyl fluoride (PMSF) was added to the assay buffer solution.
为了研究抗α2-巨球蛋白（抗A2M）抗体的结合对复合弹性蛋白酶与固相抗弹性蛋白酶抗体之间后续抗原-抗体反应的影响，将弹性蛋白酶α2-巨球蛋白复合物（EMC）与抗A2M抗体孵育，然后用固相结合的兔抗弹性蛋白酶抗体提取。EMC的两倍稀释系列产生了剂量相关的吸光度值。允许对与A2M结合的弹性蛋白酶进行免疫检测的关键因素是抗A2M抗体与溶液中的EMC预先孵育。该测定法的检测下限为每毫升0.5 ng结合的弹性蛋白酶，十名志愿者血清样本中的EMC水平显著高于血浆（28对21 ng/ml，p < 0.05，配对样本的学生t检验）。用该测定法测量的EMC水平与在测定缓冲溶液中添加苯甲基磺酰氟（PMSF）时获得的水平基本相同。
