Kravchenko I V, Furalev V A
Russian Research Center of Molecular Diagnostics and Treatment, Moscow.
Hybridoma. 1994 Feb;13(1):59-64. doi: 10.1089/hyb.1994.13.59.
Two hybridomas secreting monoclonal antibodies (mAbs) against human/rat corticotropin-releasing factor (CRF) have been produced by the cell fusion technique. Isotyping of the mAbs revealed that both belong to the IgG1 subclass. Human serum containing CRF-binding protein inhibits the binding protein inhibits the binding of CRF to both mAbs. Modification of lysine residue inhibits binding of the mAbs in a different manner. Affinity constants of binding with native and histidine-modified antigens have been determined by ELISA. The epitope specificity of the mAbs has been examined in competition experiments. No competition was detected, suggesting that the mAbs recognize different antigenic determinants. Two monoclonal antibodies can be employed in a two-site assay to measure CRF.
通过细胞融合技术制备了两种分泌抗人/大鼠促肾上腺皮质激素释放因子（CRF）单克隆抗体（mAb）的杂交瘤。单克隆抗体的亚型鉴定表明两者均属于IgG1亚类。含有CRF结合蛋白的人血清会抑制结合蛋白，进而抑制CRF与两种单克隆抗体的结合。赖氨酸残基的修饰以不同方式抑制单克隆抗体的结合。通过酶联免疫吸附测定法（ELISA）测定了与天然和组氨酸修饰抗原结合的亲和常数。在竞争实验中检测了单克隆抗体的表位特异性。未检测到竞争，这表明单克隆抗体识别不同的抗原决定簇。两种单克隆抗体可用于两点测定法来测量CRF。
