Innovation of new Florescent-epitope technique for immune assay methods

Abstract

The proposed method, which utilizes a FAM fluorescent dye and a BHQ1 quencher complex for indirect analyte assessment through antigen-antibody reactions, represents a significant advancement in immune assay technology. By leveraging the unique physical properties of the FAM-BHQ1 complex, we achieve enhanced accuracy and efficiency in analyte quantification. The complex's fluorescence characteristics, along with its inherent stability and resistance to photobleaching, contribute to improved data reproducibility. Furthermore, the system's adaptability to multiplex assays allows for simultaneous detection of multiple analytes within a single reaction, significantly increasing throughput and reducing assay time. Favorable conditions enhance the method's performance, including a broad dynamic range and minimal background signal. The inherent advantages of this approach-increased sensitivity and specificity, coupled with streamlined workflow-promise to revolutionize immunological research and diagnostics. Implementing this FAM-BHQ1-based method will facilitate more precise and reliable detection and quantification of a broader range of analytes, leading to improved diagnostic accuracy and more effective therapeutic interventions.•Despite the importance of immune assays, these assays face challenges such as standardization, variability, and issues with sensitivity.•Our propose is the use of the FAM fluorescent day, which can attach to the Fc region of the antibody and BHQ1•This limitation is the time and number of epitope-to-paratope attachments.

Keywords: BHQ-1; Epitope-paratope reaction; FAM; Florescent dye; Immunoassay; Prob antibody.

Graphical abstract

Fig. 1

The schematic illustration of the florescent- epitope detection method. A: In the presence of target analyte epitopes, it can attach to the specific antibody; therefore, the quencher-linker epitope cannot attach to the FAM antibody to suppress its light production. B: In the absence of target analyte epitopes, the BHQ1 as the quencher can attached to the FAM-antibody via a specific epitope to conquer the light emission.

Fig. 2

Physical composition of FAM-Ab plus BHQ1-Epitope. The physical possible distance between the reporter and quencher is less than 50 percent of the allowable dimension; therefore, adding a simple linker will not affect the light emission by the complex.