Terouanne B, Marchand J, Calzolari C, Monnier J, Nicolas J C, Pau B, Descomps B
Clin Chem. 1983 Feb;29(2):302-4.
Recently we developed an enzyme immunoassay involving the use of steroid delta-isomerase (EC 5.3.3.1) as enzyme label and exclusion-affinity chromatography for rapid separation of free antigen-enzyme conjugate that bound to antibodies (J. Immunol. Methods 35: 267-284, 1980). Here we describe an automated version of this procedure, for immunoassay of progesterone and choriomammotropin (human placental lactogen) in serum with the use of a centrifugal analyzer. After incubation, suitable dilutions of sera or extract plus antiserum, conjugate, and double antibody were filtered on an estradiol affinity gel-filtration column; the enzyme activity of the filtrates was determined with the centrifugal analyzer. Results correlate well with those obtained by radioimmunoassay: r (progesterone) = 0.980, r (choriomammotropin) = 0.940. The within-run and between-run precision, specificity, sensitivity, accuracy, and speed of this system make it a useful tool for immunoassay.
最近,我们开发了一种酶免疫测定法,该方法使用类固醇δ-异构酶(EC 5.3.3.1)作为酶标记,并采用排阻亲和色谱法快速分离与抗体结合的游离抗原-酶缀合物(《免疫学方法杂志》35:267 - 284,1980年)。在此,我们描述了该方法的自动化版本,用于使用离心分析仪测定血清中的孕酮和绒毛膜促乳腺素(人胎盘催乳素)。温育后,将血清或提取物加上抗血清、缀合物和双抗体的适当稀释液在雌二醇亲和凝胶过滤柱上进行过滤;用离心分析仪测定滤液的酶活性。结果与放射免疫测定法获得的结果相关性良好:r(孕酮) = 0.980,r(绒毛膜促乳腺素) = 0.940。该系统的批内和批间精密度、特异性、灵敏度、准确性和速度使其成为免疫测定的有用工具。