Goldstein A M, Marcon L, Cullen B R, Nelson D L
Immunophysiology Section, National Cancer Institute, Bethesda, MD 20892.
J Immunol Methods. 1988 Feb 24;107(1):103-9. doi: 10.1016/0022-1759(88)90015-4.
A solid-phase, competition enzyme-linked immunosorbent assay (ELISA) was established for the quantitative measurement of soluble (human) interleukin-2 receptors (IL-2R). The ladder of reagents from the solid phase up consisted of: (1) recombinant DNA-derived, purified IL-2R, (2) sample-containing soluble IL-2R and fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody, 7G7/B6, directed against the IL-2R, (3) alkaline phosphatase-conjugated rabbit anti-FITC, and (4) substrate. This ELISA was compared with a 'sandwich' ELISA for soluble IL-2R. The competitive ELISA was less sensitive than the 'sandwich' assay, being capable of measuring 5000 versus 31 U/ml, respectively. While both anti-Tac and 7G7/B6 in the IL-2R-containing sample inhibited the 'sandwich' assay, only 7G7/B6 inhibited the competition assay. Anti-mouse immunoglobulin enhanced the 'sandwich' assay and inhibited the competitive assay; both effects could be overcome by the addition of normal mouse immunoglobulin in the sample buffer. Studies of a patient's serum receiving anti-Tac as therapy for the adult T cell leukemia demonstrated that rises in the level of IL-2R occurring with anti-Tac therapy, as measured with the competition assay, were masked in the 'sandwich' assay. This competition ELISA will be useful for measuring soluble IL-2R levels in patients receiving anti-Tac as therapy for various immunologic disorders.
建立了一种固相竞争酶联免疫吸附测定法（ELISA），用于定量检测可溶性（人）白细胞介素-2受体（IL-2R）。从固相开始的试剂梯级包括：（1）重组DNA衍生的纯化IL-2R，（2）含样品的可溶性IL-2R和针对IL-2R的异硫氰酸荧光素（FITC）偶联单克隆抗体7G7/B6，（3）碱性磷酸酶偶联的兔抗FITC，以及（4）底物。将这种ELISA与用于可溶性IL-2R的“夹心”ELISA进行比较。竞争ELISA的灵敏度低于“夹心”测定法，分别能够检测5000和31 U/ml。虽然含IL-2R样品中的抗Tac和7G7/B6都抑制“夹心”测定法，但只有7G7/B6抑制竞争测定法。抗小鼠免疫球蛋白增强“夹心”测定法并抑制竞争测定法；在样品缓冲液中加入正常小鼠免疫球蛋白可以克服这两种效应。对一名接受抗Tac治疗成人T细胞白血病的患者血清进行的研究表明，用竞争测定法测量时，抗Tac治疗期间出现的IL-2R水平升高在“夹心”测定法中被掩盖。这种竞争ELISA将有助于测量接受抗Tac治疗各种免疫疾病患者的可溶性IL-2R水平。
