Thermodynamic and kinetic approaches for evaluation of monoclonal antibody - Lipoprotein interactions

Abstract

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.

Capillary Electrophoresis: Trends and Recent Advances

“… 86 Partial filling (PF) ACE was combined with adsorption energy distribution to determine the heterogeneity of interaction of apoB-100 containing lipoproteins and their antibodies. 87 The interaction proved homogeneous and PF-ACE results were in alignment with quartz crystal microbalance experiments.…”

Correlation between Cholesterol, Triglycerides, Calculated, and Measured Lipoproteins: Whether Calculated Small Density Lipoprotein Fraction Predicts Cardiovascular Risks

“…These lipoproteins are actually mixtures of various proportions of esterified and nonesterified cholesterol, phospholipids, proteins, triglycerides, and surface apolipoproteins [ 8 ]. Kinetic studies have identified a lot of variability in terms of shape, size, and lipid composition which are difficult to measure as perfection in clinical laboratories provided improvement in laboratory science and calibration practices [ 9 ]. The recent data has subcategorized LDL particles based upon their size and density into small and dense LDL-cholesterol particles termed small density LDLc (sdLDLc) and large dense LDL-cholesterol, which has been proven to be more predictive to highlight underlying cardiovascular risks [ 10 ].…”

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“… 1 However, for detailed characterizations of interactions between more complicated and larger biomolecules, for example, biological drugs and their target receptors in biochemical assays, more advanced data analysis approaches are crucial. 3 − 5 These approaches should not be considered a replacement for carefully executed experiments but as a way to deal with situations where more complicated interactions are present.…”