浅表带蛋白特异性跨种单克隆抗体检测人和动物体液中浅表带蛋白。

J. Su, B. Schumacher, K. M. Lindley, V. Soloveychik, W. Burkhart, J. Triantafillou, K. Kuettner, T. Schmid
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引用次数: 88

摘要

在本报告中,我们描述了人浅带蛋白(SZP)的纯化,跨物种单克隆抗体(mab)的产生和该蛋白在人和动物体液中的检测。作为免疫抗原的人SZPs,是从人软骨器官培养基或人滑膜液中纯化出来的。免疫抗原与RIBI佐剂以三种形式之一混合:未修饰的SZP,浅区蛋白-锁孔帽贝血青素偶联物(SZP- klh),或浅区蛋白和透明质酸的混合物(SZP- ha)。生成了包括GW4.23、S6.79、S13.52、S13.233和S17.109在内的单克隆抗体,并对其进行了表征。单克隆抗体(MAb) S6.79是一种IgG2b,从SZP-KLH免疫中获得K(D) 3.14 x 10(-9) M,特别令人感兴趣。在酶联免疫吸附试验(ELISA)和Western blotting中,它对大分子量的SZP有强烈反应。免疫组化染色最浅层的关节软骨,而软骨的中间和深层不染色。当MAb S6.79应用于体液的Western blots时,在滑液样品中检测到一个强345-kDa条带,在血浆和血清样品中检测到类似大小的较弱条带。经Western blotting和抗体吸收实验证实,MAb S6.79在牛、狗、豚鼠和兔的滑液样品中也显示出与SZP的跨物种免疫反应性。这种跨物种单抗将成为人类和动物模型研究中监测SZP水平和组织分布的有用工具。它可能有助于确定SZP在正常关节中的作用,并可能对骨关节炎、类风湿关节炎和campto拇趾-关节病-髋外翻-心包炎等病理条件下测量SZP具有诊断或预后价值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Detection of superficial zone protein in human and animal body fluids by cross-species monoclonal antibodies specific to superficial zone protein.
In this report we describe the purification of human superficial zone protein (SZP), the generation of cross-species monoclonal antibodies (MAbs) and the detection of this protein in human and animal body fluids. Human SZPs, used as immunizing antigens, were purified either from culture media of human cartilage organ cultures or from human synovial fluids. The immunizing antigens were mixed with RIBI adjuvant in one of three forms: nonmodified SZP, superficial zone protein-keyhole limpet hemocyanin conjugate (SZP-KLH), or a mixture of superficial zone protein and hyaluronic acid (SZP-HA). A panel of MAbs including GW4.23, S6.79, S13.52, S13.233, and S17.109 were generated and characterized. Monoclonal antibody (MAb) S6.79, an IgG2b with K(D) 3.14 x 10(-9) M from SZP-KLH immunization, is of particular interest. It reacts strongly to a large molecular weight form of SZP in both enzyme-linked immunosorbent assay (ELISA) and Western blotting. It stains the most superficial layer of articular cartilage in immunohistochemistry, whereas the middle and deep zones of cartilage are not stained. When MAb S6.79 was applied to Western blots of human body fluids, a strong 345-kDa band was detected in samples of synovial fluid and weaker bands of similar size were detected in samples of plasma and serum. MAb S6.79 also showed cross-species immunoreactivity with SZP in samples of synovial fluids harvested from bovine, dog, guinea pig, and rabbit, as demonstrated by Western blotting and antibody absorption experiments. This cross-species MAb will be a useful tool in human and animal model studies for monitoring SZP levels and tissue distribution. It may help define the roles of SZP in normal articular joints and may be of diagnostic or prognostic value for the measurement of SZP in pathological conditions such as osteoarthritis, rheumatoid arthritis, and camptodactyly-arthropathy-coxa vara-pericarditis.
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来源期刊
Hybridoma
Hybridoma 医学-免疫学
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