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Detection of antigen-antibody affinity by fluorescence autocorrelation spectroscopy

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Detection of antigen-antibody affinity by fluorescence autocorrelation spectroscopy

[Abstract]:
In this paper, the specific realization and potential application of fluorescence autocorrelation spectroscopy (FACS) technology as an immunoassay technology were explored. In the experiment, the interaction between Alexa Fluor647 fluorescent molecule and green fluorescent protein antigen and antibody was used as the model, and a desktop fluorescence correlation spectrometer was used to collect the fluorescence autocorrelation data of fluorescent molecular antigen and antibody mixture of different concentrations. Combining the traditional FACS technique with the maximum entropy method, we can quantitatively and qualitatively evaluate the degree of antigen-antibody binding. The acquired fluorescence autocorrelation data were analyzed using self-developed data analysis software to obtain antigen-antibody dissociation constants. The experimental results show that the FACS technology applied to antigen-antibody affinity detection has the characteristics of simplicity, rapidity and sensitivity; the detection limit is between picomolar and nanomolar; the modular data analysis software can standardize the operation process and improve the accuracy of data analysis. and repeatability.