Brandt E, Altman A, Grünefeld M, Ulmer A J, Flad H D
Immunobiology. 1986 Aug;172(1-2):33-53. doi: 10.1016/S0171-2985(86)80051-1.
Human recombinant interleukin 2 (r-IL2) was used as an immunizing antigen to yield a murine monoclonal antibody (mAb) termed BO-7. Although the antibody binds to r-IL2 more avidly, it also reacted strongly with IL2 from natural sources in an enzyme-linked immunosorbent assay (ELISA), allowing the detection of the purified lymphokine at sensitivity levels closely approaching those found with the IL2 biological assay. Binding to the antigen is specific, as deduced from the close correlation of ELISA immunoreactivity with IL2 biological activity and from immunoblot analysis of electrophoretically separated IL2 from various sources. Binding studies with synthetic IL2-derived peptides revealed the location of the epitope, which is recognized by mAb BO-7: A peptide representing amino acid residues 59-72 (peptide 84) is strongly reactive with the antibody, while an overlapping peptide (residues 48-69) is not. Peptide 84, moreover, can be applied for immunopurification of mAb BO-7 and competes for binding to the antibody with the intact IL2 molecule. In turn, another monoclonal anti-IL2 antibody (35H10), showing the same reactivity pattern with peptides, competes with mAb BO-7 for binding to IL2. The application of mAb BO-7 as a specific reagent for the quantitation of IL2 in a sandwich-type ELISA is demonstrated.
人重组白细胞介素2（r-IL2）用作免疫抗原，以产生一种名为BO-7的鼠单克隆抗体（mAb）。尽管该抗体与r-IL2的结合更具亲和力，但在酶联免疫吸附测定（ELISA）中它也与天然来源的IL2发生强烈反应，从而能够以接近IL2生物学测定的灵敏度水平检测纯化的淋巴因子。从ELISA免疫反应性与IL2生物学活性的密切相关性以及对来自各种来源的电泳分离的IL2进行免疫印迹分析可以推断，与抗原的结合是特异性的。与合成的IL2衍生肽的结合研究揭示了被mAb BO-7识别的表位的位置：代表氨基酸残基59-72的肽（肽84）与该抗体强烈反应，而重叠肽（残基48-69）则不反应。此外，肽84可用于mAb BO-7的免疫纯化，并与完整的IL2分子竞争与抗体的结合。反过来，另一种单克隆抗IL2抗体（35H10）与肽显示相同的反应模式，它与mAb BO-7竞争与IL2的结合。证明了mAb BO-7作为夹心型ELISA中定量IL2的特异性试剂的应用。
