Wada S, Naylor P H, Naylor C W, Goldstein A L
Department of Biochemistry, George Washington University School of Medicine and Health Sciences, Washington, DC 20037.
Int J Immunopharmacol. 1988;10(7):795-801. doi: 10.1016/0192-0561(88)90002-1.
An improved microELISA to measure thymosin alpha 1 (T alpha 1) is described which uses a rabbit antibody against T alpha 1 that has been absorbed with a synthetic C-14 fragment of T alpha 1. This assay is compared to the previous assay which used the whole antisera. The antibodies to T alpha 1 are preincubated with the standard or human sera overnight at 4 degrees C, then incubated for an additional 24 h in microtiter plates coated with T alpha 1. Using the whole antiserum, the average T alpha 1 level was 2480 +/- 1110 (mean +/- S.D.) pg/ml by ELISA and 2360 +/- 870 pg/ml by radioimmunoassay (RIA) in eight different samples of human cord sera. Using the N-specific absorbed antiserum the mean T alpha 1 level was 11,800 +/- 4800 pg/ml by ELISA and 10,600 +/- 5200 pg/ml by RIA. Recoveries of exogenously added T alpha 1 are complete (109 +/- 25% for whole and 108 +/- 15% for absorbed antisera). The absorbed antiserum has an increased affinity for the amino acid terminal region of T alpha 1 and the T alpha 1 values by use of absorbed antisera are significantly higher (3-5 x) than those measured using the whole antisera. Thus, the absorbed antisera produces an ELISA which is more sensitive and specific for serum thymosin alpha 1.
本文描述了一种改进的微量酶联免疫吸附测定法（microELISA），用于检测胸腺素α1（Tα1）。该方法使用针对Tα1的兔抗体，该抗体已用Tα1的合成C-14片段进行吸附处理。将此测定法与先前使用全抗血清的测定法进行比较。将Tα1抗体与标准品或人血清在4℃下预孵育过夜，然后在包被有Tα1的微量滴定板中再孵育24小时。使用全抗血清时，在八份不同的人脐带血清样本中，通过酶联免疫吸附测定法测得的平均Tα1水平为2480±1110（均值±标准差）pg/ml，通过放射免疫测定法（RIA）测得的平均Tα1水平为2360±870 pg/ml。使用N特异性吸附抗血清时，通过酶联免疫吸附测定法测得的平均Tα1水平为11,800±4800 pg/ml，通过放射免疫测定法测得的平均Tα1水平为10,600±5200 pg/ml。外源添加的Tα1回收率完全（全抗血清为109±25%，吸附抗血清为108±15%）。吸附抗血清对Tα1的氨基酸末端区域具有更高的亲和力，使用吸附抗血清测得的Tα1值比使用全抗血清测得的值显著更高（3 - 5倍）。因此，吸附抗血清产生的酶联免疫吸附测定法对血清胸腺素α1更敏感且更具特异性。
