A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5′-diphosphocholine coupled dendrimers
A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5′-diphosphocholine coupled dendrimers
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DOI：
10.1016/j.jim.2009.02.002
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发表时间：
2009-04-15
影响因子：
2.2
通讯作者：
Boas, Ulrik
中科院分区：
文献类型：
--
作者：
Heegaard, Peter M. H.;Pedersen, Heidi G.;Boas, Ulrik
C-反应蛋白(CRP)是一种重要的急性时相蛋白，被用作炎症和感染的敏感指标，也与心血管问题的风险有关。基于C反应蛋白与磷胆碱的亲和力，本文建立了一种稳健而灵敏的检测C反应蛋白的酶联免疫吸附试验。在这个设计中，合成的球状聚合物(树枝状大分子)被用作多价展示磷胆碱分子的支架。存在于样本中的C反应蛋白与涂层中密度较高的磷胆碱部分结合，并可被特定抗体检测到。用市售的抗人CRP.抗体对猪和人的CRP进行了检测，结果表明，该方法比以往报道的白蛋白偶联胞苷二磷胆碱免疫分析法更灵敏。包被物在室温下至少稳定30天，批内和批间重复性好。将结果与免疫比浊法和商业ELISA试剂盒进行比较，结果与免疫比浊法非常一致，但与商业测定不一致，可能是由于校准差异所致。通过提供具有所需物种特异性的足够的检测抗体，该方法也适用于其他物种。(C)2009爱思唯尔B.V.保留所有权利。
C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP.The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity. (C) 2009 Elsevier B.V. All rights reserved.