Complement factor H as a marker for detection of bladder cancer
Complement factor H as a marker for detection of bladder cancer
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DOI：
10.1373/clinchem.2004.042192
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发表时间：
2005-05-01
影响因子：
9.3
通讯作者：
Jokiranta, TS
中科院分区：
文献类型：
--
作者：
Cheng, ZZ;Corey, MJ;Jokiranta, TS
背景资料：用于膀胱癌的BTA TRAK（TM）和BTA stat（TM）检测使用单克隆抗体（mAb）X13.2和X52.1来检测尿液中的H因子（FH）相关物质。方法：采用Western blot分析纯化的FH、重组H因子相关蛋白1（FHR-1）以及血清和尿液样品，以鉴定X13.2和X52.1的配体。重组FH构建体用于鉴定X13.2和X52.1的靶位点。为了分析FH的天然配体是否可以与捕获mAb X52.1竞争其识别，我们使用表面等离子体共振分析。结果：X13.2与FH的结构域3和FH样蛋白1结合，而X52.1与FH的结构域18和FHR-1结合。使用从膀胱癌患者的尿液和纯化的FH的特定FH消耗，我们证明了FH是由BTA TRAK测试识别的配体。相反，尿中的FHR-1降低了FH依赖的检测信号。结论：FH是膀胱癌的肿瘤标志物。为了揭示膀胱癌的存在，BTA TRAK测定检测FH，而FHR-1能够部分抑制这种检测。这表明基于单个样品中同时阳性和阴性信号的组合效应的诊断免疫测定的特殊机制。(c)2005年美国临床化学协会。
Background: The BTA TRAK (TM) and BTA stat (TM) tests for bladder cancer use monoclonal antibodies (mAbs) X13.2 and X52.1 to detect factor H (FH)-related material in urine. The exact ligands remain unknown.Methods: Western blot analyses of purified FH, recombinant factor H-related protein 1 (FHR-1), and serum and urine samples were used to identify the ligands of X13.2 and X52.1. Recombinant FH constructs were used to identify the target sites of X13.2 and X52.1. To analyze whether natural ligands of FH could compete with its recognition by the capture mAb X52.1, we used surface plasmon resonance analysis. The role of the ligands of X52.1 in the BTA TRAK assay was tested with use of purified proteins and FH-depleted samples.Results: X13.2 bound to domain 3 of FH and FH-like protein 1, whereas X52.1 bound to domain 18 of FH and to FHR-1. Using specific FH depletion from a bladder cancer patient's urine and purified FH, we demonstrated that FH is the ligand recognized by the BTA TRAK test. By contrast, FHR-1 in urine reduced the FH-dependent test signal.Conclusions: FH is a tumor marker for bladder cancer. To reveal the presence of bladder cancer, the BTA TRAK assay detects FH, whereas FHR-1 is able to partly inhibit this detection. This indicates a special mechanism for a diagnostic immunoassay based on the combined effect of simultaneous positive and negative signals in a single sample. (c) 2005 American Association for Clinical Chemistry.