Antigen-Adjuvant Interactions in Vaccines by Taylor Dispersion Analysis: Size Characterization and Binding Parameters

Abstract

Vaccine adjuvants are immunostimulatory substances used to improve and modulate the immune response induced by antigens. A better understanding of the antigenadjuvant interactions is necessary to develop future effective vaccine. In this study, Taylor dispersion analysis (TDA) was successfully implemented to characterize the interactions between a polymeric adjuvant (poly(acrylic acid), SPA09) and a vaccine antigen in development for the treatment of Staphylococcus aureus. TDA allowed to rapidly determine both (i) the size of the antigen-adjuvant complexes under physiological conditions, and (ii) the percentage of free antigen in the adjuvant/antigen mixture at equilibrium, and finally get the interaction parameters (stoichiometry and binding constant). The complex sizes obtained by TDA were compared to the results obtained by transmission electron microscopy (TEM) and the binding parameters were compared to results previously obtained by frontal analysis continuous capillary electrophoresis (FACCE).

Characterization of antigen adjuvant interactions in polyacrylate adjuvanted vaccines

“…Alternative methods can also be used to characterize antigen-polyacrylate formulations. For instance, Malburet et al have used capillary electrophoresis to study the interaction of SPA09 with PrsA, as mentioned before [ 11 , 36 ]. Vialle et al have used a competitive ELISA technique to study the interaction of a polyacrylate gel adjuvant, Montanide Gel™, with model proteins [ 27 ].…”

β‐Cyclodextrin and folic acid host–guest interaction binding parameters determined by Taylor dispersion analysis and affinity capillary electrophoresis

“…Taylor dispersion analysis (TDA) is a flow-based approach used for assessing diffusion coefficient (D) and hydrodynamic radius (R h ) of the analyte with sizes ranging from small molecules, peptides, proteins to nanoparticles [36][37][38]. Dispersion of an analyte plug occurs in a laminar Poiseuille flow in accordance with the TDA theory, which states that convection (axial direction) and molecular (radial direction) diffusion work together to disperse an analyte plug [39,40].…”

“…CE-based TDA was first used to study the protein-ligand interaction at equilibrium and simultaneously characterize the size of the protein [43]. TDA was recently shown to be an effective and general method for finding equilibrium binding parameters for biomolecules [38,44].…”

3D Printed Instrument for Taylor Dispersion Analysis with Two-Point Laser-Induced Fluorescence Detection

“…TDA has evolved from studying gaseous diffusion coefficients in large tubes to capillary-based solution phase measurements of small molecules, , macromolecular complexes, − and even micron-scale particles . In TDA, analysis time is proportional to the square of the tubing radius.…”