Bashir I, Sikora K, Foster C S
Department of Cellular and Molecular Pathology, University of Liverpool, UK.
Virchows Arch. 1998 Mar;432(3):279-87. doi: 10.1007/s004280050166.
A novel immunization protocol together with stringent selection criteria have been employed to generate a new murine monoclonal antibody ("D8", isotype IgG1, kappa) which specifically recognizes the human p170 drug resistance glycoprotein. This antibody is directed towards a defined peptide sequence located in the -COOH terminal region of the first external loop of the molecule. It is reactive with its epitope within the intact native glycoprotein in formalin-fixed and conventionally processed histological tissues, in flow-cytometric preparations and by Western blotting. The antibody precipitates its target peptide sequence from solution, and thus may be a useful reagent with which to establish an ELISA, RIMA or other similar assay. The peptide epitope recognized by this monoclonal antibody is restricted to the human MDR1 gene product and is not contained within the rodent homologue of the P-170 molecule. Immunohistochemistry has consistently failed to detect this epitope in rodent tissues, thus confirming that it does not exhibit the cross-reactivity of other currently available anti-P-glycoprotein monoclonal antibodies. The experience of this study emphasizes the value of the tuberculin-PPD (purified protein derivative) immunization protocol as a powerful strategy when generating monoclonal antibodies to small synthetic peptides. The resulting monoclonal antibody (D8) will be an invaluable reagent with which to analyse P-170 glycoprotein expression when assessing the role of multidrug resistance in human cancers.
一种新的免疫方案与严格的筛选标准相结合，已用于产生一种新的鼠单克隆抗体（“D8”，同种型IgG1，κ链），该抗体特异性识别人类p170耐药糖蛋白。这种抗体针对位于该分子第一个外环的-COOH末端区域的特定肽序列。它能与福尔马林固定和常规处理的组织学组织、流式细胞术制备物以及蛋白质免疫印迹中的完整天然糖蛋白内的表位发生反应。该抗体能从溶液中沉淀出其靶肽序列，因此可能是建立酶联免疫吸附测定（ELISA）、放射免疫测定（RIMA）或其他类似检测方法的有用试剂。这种单克隆抗体识别的肽表位仅限于人类多药耐药基因1（MDR1）基因产物，不包含在P - 170分子的啮齿动物同源物中。免疫组织化学一直未能在啮齿动物组织中检测到该表位，从而证实它不表现出其他现有抗P - 糖蛋白单克隆抗体的交叉反应性。本研究的经验强调了结核菌素 - 纯化蛋白衍生物（PPD）免疫方案作为一种强大策略在产生针对小合成肽的单克隆抗体时的价值。所得的单克隆抗体（D8）在评估多药耐药性在人类癌症中的作用时，将是分析P - 170糖蛋白表达的宝贵试剂。
