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QUANTITATIVE STUDY OF ANTIGEN-ANTIBODY INTERACTION USING FLUORESCENCE CORRELATION SPECTROSCOPY
- Categories:FCS Technology for Molecular Interaction Analysis
- Time of issue:2023-05-24 14:28
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(Summary description)The team of Sergey Y. Tetin from Abbott Laboratories, USA, reviewed the application research of fluorescence correlation spectroscopy ( FCS ) and its derivative techniques in the quantitative analysis of antigen - antibody binding reactions. Antibody molecules are widely used in fields such as basic research and medical testing. Therefore, analytical techniques that can quantitatively and homogeneously analyze antigen - antibody interactions in solution samples are of great academic and commercial value. Traditional biological techniques (such as ELISA ) are difficult to quickly, sensitively, and accurately characterize the antigen - antibody binding reaction in a small amount (several microliters) of samples , including the equilibrium constant, kinetic constant, and antigen-specific binding site of the reaction. , Antigen - antibody binding model, etc. Fluorescence autocorrelation ( FACS ) and cross-correlation ( FCCS ) spectroscopic techniques can quantitatively analyze various characteristics of antigen - antibody binding reactions in trace solutions without separation (homogeneous phase) of antigen - antibody conjugates. FACS technology can be applied to determine the equilibrium dissociation constant of antigen - antibody binding reaction, and determine the key binding site on the antigen molecule ; FCCS technology can be combined with the sandwich method to determine the antigen- Kinetic dissociation constant of antibody binding and binding reaction ; moment analysis technique ( Moment analysis ) can be used for antibody binding activity analysis ; time-integrated fluorescence accumulation analysis ( TIFCA ) can establish antigen - antibody binding model . In summary, the application of FCS and its derivative technology to the study of antigen - antibody interaction has the advantages of high sensitivity, less sample consumption, rapid and homogeneous detection, etc., which is of great significance for the development of antibody drugs
QUANTITATIVE STUDY OF ANTIGEN-ANTIBODY INTERACTION USING FLUORESCENCE CORRELATION SPECTROSCOPY
- Categories:FCS Technology for Molecular Interaction Analysis
- Time of issue:2023-05-24 14:28
- Views:
Abstract
Antibodies are excellent binding proteins that have found numerous applications in biological research, biotechnology, and medicine. Characterization of their ligand binding properties has long been, and continues to be, the focus of many researchers. Antibodies are also perfect test systems which can be used for the evaluation of newly introduced biophysical techniques. Working with many different antibodies, we continuously implement the growing arsenal of methods offered by fluorescence fluctuation spectroscopy (FFS) and apply them for antibody research. In this chapter, we will describe applications of FFS for antibody binding characterization and also provide examples how studying of antibodies helps to develop and enhance the tool set offered by FFS technology. In addition to traditional affinity evaluations, we will describe how resolving molecular populations enables determinations of the binding stoichiometry and provides further information about the system. Even though all our examples include antibodies, the same experimental procedures can also serve well for characterizing v arious proteins and other ligand binding systems.
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