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Published on: July 13, 2016
The quantitative detection method employing a combination of high-affinity antibodies targeting PCT
Xiaoxia Cheng1,2, Lichen Zha1,2, Jiao Yang2,3
1Department of Clinical Laboratory, Suzhou Hospital, Affiliated Hospital of Medical School, Nanjing University, Suzhou, Jiangsu, China.
Background And Aims:
Procalcitonin (PCT) is a widely recognized inflammation marker utilized in various clinical testing contexts and is subject to ongoing refinements, thereby imposing greater demands on core antibodies. However, the published literature lacks a comprehensive description of them.
Material And Methods:
In this study, we initially expressed the full-length PCT protein in eukaryotic systems, followed by conventional antibody engineering techniques for in vitro cell fusion and the selection of positive hybridoma cell clones specific to the PCT protein.
Results:
A total of 83 positive clones were generated, among which 15 high-affinity IgG1 subtype monoclonal antibodies were selected for complementarity-determining region (CDR) analysis to predict potential antigen-recognition epitopes. We provided a detailed characterization of the binding properties between these high-affinity antibodies and the PCT protein. Utilizing time-resolved fluorescent microsphere (TRFM), a novel fluorescent microsphere-based immunochromatographic strip (FM-ICS) approach was developed, with MomAb-8 serving as the capture antibody and MomAb-20 functioning as the labeling antibody. Ultimately, following the preliminary evaluation of clinical samples, it was demonstrated that this FM-ICS exhibited a favorable linear range, stability, and clinical relevance.
Conclusion:
This study presents a novel approach to enhancing the efficiency of antibody screening across a diverse array of combinations. Furthermore, the method established herein holds significant potential for clinical application in detecting PCT protein using FM-ICS.
