Development, identification and application of 33 monoclonal antibodies against cardiac troponin T.

Yuehong HU; Zimin Chen; Yuxiang Chen; Yinghui Yang; Shuying Wei; Liuwei Song; Guoliang Zhou; Shengxiang GE

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Development, identification and application of 33 monoclonal antibodies against cardiac troponin T.

Author: Yuehong HU ¹ ; Zimin CHEN ² ; Yuxiang CHEN ¹ ; Yinghui YANG ¹ ; Shuying WEI ² ; Liuwei SONG ² ; Guoliang ZHOU ² ; Shengxiang GE ²
Author Information

1. Laboratory of Lung, Xinglin Branch of the First Affiliated Hospital of Xiamen University, Xiamen 361022, Fujian, China.
2. National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian, China.
Publication Type:Journal Article
Keywords: acute myocardial infarction (AMI); cardiac troponin T; chemiluminescence; monoclonal antibodies
MeSH: Antibodies, Monoclonal; immunology; Enzyme-Linked Immunosorbent Assay; Epitopes; immunology; Humans; Hybridomas; Luminescent Measurements; Myocardial Infarction; Peptide Fragments; Sensitivity and Specificity; Troponin T; immunology
From: Chinese Journal of Biotechnology 2016;32(12):1694-1703
CountryChina
Language:Chinese
Abstract: The aim of this study is to prepare and characterize cardiac troponin T (cTnT) monoclonal antibodies (mAb), and further develop a chemiluminescence quantitative detection assay for cTnT. BALB/c mice were immunized with recombinant cTnT antigen, and specific mAbs were prepared using conventional hybridoma technique and screened by indirect ELISA method. To identify the epitopes, several cTnT peptide fragments were synthesized or expressed by genetic engineering. A double antibody sandwich ELISA method was used to screen the mAb pairs for cTnT detection, and the automatic chemiluminescence detection assay for cTnT was developed. In total 220 clinical specimens were used for system comparison between our assay and Roche cTnT assay; further performance characteristics was evaluated by testing 238 clinical samples and 784 physical examination samples. We successfully screened 33 strains of hybridoms against cTnT, and the mAbs' epitopes were identified. Mab E16H8 and C8G11 with a detection limit of 10 pg/mL cTnT antigen were selected to develop the full automatic chemiluminescence quantitative assay. The correlation coefficient of our reagent with Roche's was 0.959 9, with a coincidence rate of 95%. The assay presented a sensitivity of 97.5%, and a specificity of 99.15% in detection of clinical samples. The cTnT concentration was less than 0.080 6 ng/mL in 99% of general population, which agrees with the definition of WHO on patients with acute myocardial infarction (AMI). In summary, we developed monoclonal antibodies against predominant epitopes for diagnostics of cTnT, and an automatic tubular chemiluminescence quantitative detection assay was further developed, which presents a high coincidence rate with Roche's.