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

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• 作者：Evgen Multia^a ; Heli Siré ; n^a ; Karl Andersson^b ; ^c ; Jö ; rgen Samuelsson^d ; Patrik Forssé ; n^d ; Torgny Fornstedt^d ; Katariina Ö ; ö ; rni^e ; Matti Jauhiainen^f ; Marja-Liisa Riekkola^a ; ^{marja-liisa.riekkola@helsinki.fi}
• 关键词：Monoclonal anti-apoB-100 antibody ; Lipoproteins ; Thermodynamics ; Kinetics ; Interaction map
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：1 February 2017
• 年：2017
• 卷：518
• 期：Complete
• 页码：25-34
• 全文大小：1894 K
• 卷排序：518

文摘

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.