Chemiluminescent immunoassay for high-sensitivity C-reactive protein.
- Author:
Pingguo LIU
1
;
Guoqiang LI
;
Yixin CHEN
;
Haifeng LUO
;
Dedang HUANG
;
Yingbin WANG
;
Shengxiang GE
;
Jun ZHANG
;
Ningshao XIA
Author Information
1. Department of Hepatobiliary Surgery, Zhongshan Hospital, Xiamen University, Xiamen 361004, China.
- Publication Type:Journal Article
- MeSH:
C-Reactive Protein;
analysis;
chemistry;
Humans;
Immunoassay;
methods;
Luminescent Measurements;
methods;
Sensitivity and Specificity
- From:
Chinese Journal of Biotechnology
2010;26(8):1150-1156
- CountryChina
- Language:Chinese
-
Abstract:
We developed a high-sensitivity C-reactive protein quantifiable chemiluminescent immunoassay (hs-CRP CLIA). The high-purity native CRP was purified from hepatic cirrhosis patient ascetic fluid by affinity and ion exchange chromatography and used as an immunogen to develop the monoclonal antibodies (mAbs) against CRP. Twenty-two mAbs were identified reactive with CRP in ELISA and 13 of them were reactive in the phosphorycholine ligand capture ELISA. The mAbs 10C5 and 10C11 were selected to develop the hs-CRP CLIA. The linearity and performance of the hs-CRP CLIA was characterized. It was showed not reactive when testing against other serum materials (IgG, hemoglobin and triglyceride). The reliable correlation (R2 > 0.993) was obtained between testing value (RLU/S) and the concentration of human serum CRP calibrator. The linearity fell in the range of 0.04-20.38 mg/L. The assay has good accuracy and reproducibility, the mean recovery was 99% and the precision of the intra- and inter assay was CVs (4.2%-5.8%) and (9.0%-11.5%), respectively. In testing of 90 human sera, this assay performed well and correlated comparably with a commercial hs-CRP ELISA kit. Thus, hs-CRP CLIA is an accurate, reliable, quantifiable assay for detection of high-sensitive C-reactive protein in serum, it may be useful to improve the risk assessment of cardiovascular disease and the prognosis of inflammatory bowel disease.
