Thermodynamic and kinetic approaches for evaluation of monoclonal antibody Lipoprotein interactions.
Two complementary instrumental techniques were used, and the data generated was processed with advanced numerical tools to investigate the interactions between anti human apoB 100 monoclonal antibody (anti apoB 100 Mab) and apoB 100 containing lipoproteins. Partial Filling Affinity Capillary Electrophoresis (PF ACE) combined with Adsorption Energy Distribution (AED) calculations provided information on the heterogeneity of the interactions without any a priori model assumptions. The AED calculations evidenced a homogenous binding site distribution for the interactions. Quartz Crystal Microbalance (QCM) studies were used to evaluate thermodynamics and kinetics of the Low Density Lipoprotein (LDL) and anti apoB 100 Mab interactions. High affinity and selectivity were observed, and the emerging data sets were analysed with so called Interaction Maps. In thermodynamic studies, the interaction between LDL and anti apoB 100 Mab was found to be predominantly enthalpy driven. Both techniques were also used to study antibody interactions with Intermediate Density (IDL) and Very Low Density (VLDL) Lipoproteins. By screening affinity constants for IDL VLDL sample in a single injection we were able to distinguish affinity constants for both subpopulations using the numerical Interaction Map tool.
Analytical biochemistry