Thermodynamic analysis of the interactions of a mouse dinitrophenyl‐specific myeloma protein, MOPC 315, with immobilized dinitrophenyl and trinitrophenyl ligands by affinity electrophoresis

Abstract

Thermodynamic analysis of the interactions of a mouse dinitrophenyl-specific myeloma protein, MOPC 3 15, with immobilized dinitrophenyl and trinitrophenyl ligands by affrnity electrophoresis* Affinity electrophoresis was applied to determine the dissociation constants and the thermodynamic parameters for the interactions between the mouse myeloma, MOPC 3 15, and trinitrophenyl (Tnp) and dinitrophenyl (Dnp) haptens. The myeloma protein consists of monomeric, dimeric, and trimeric forms of IgA. The dimer had a higher affinity to the haptens than the monomer, but its Fab' fragment had lower affinity than the monomer. All of these myeloma proteins had higher affinities to Tnp-hapten than to Dnphapten. The affinity of MOPC 3 15 proteins increased when the temperature increased. The van't Hoff plots for all ofthese interactions gave straight lines within the temperature range from 7 "C to 40 "C. The hN" were calculated by using the linear van? Hoff plots. The AS" were calculated from the AW and AGO. The AH0 and ASo were both positive. Hence, the interactions were endothermic and hydrophobic.

Polyvalent Interactions in Biological Systems: Implications for Design and Use of Multivalent Ligands and Inhibitors

“…The first example is the binding of a bivalent antibody to ligands that are densely packed on a biological surfaceÐsuch as a mammalian cell, a virus, or a solid support for an enzyme-linked immunosorbant assay (ELISA)Ðor immobilized in a polymeric matrix ( Figure 8). [276] In general, monovalent binding constants of antibodies for small organic ligands vary significantly, but are in the range of 10 5 ± 10 8 m À1 . For non-or positively cooperative systems, we expect K bi 2 !…”

Observations on the Binding of Four Anti-carbohydrate Monoclonal Antibodies to Their Homologous Ligands

“…Indeed, in a case where both hydrogen bonding and significant hydrophobic interactions had been found by x-ray diffraction analysis of the antibody having the immunodeterminant bound to its binding site (Cygler et al, 1991), it has been shown that antigenantibody interactions can also have favorable entropy contributions (Sigurskjold and Bundle, 1992) to the binding. In another case, thermodynamic van't Hoff analysis of the interaction between di-and trinitrophenyl ligands with monoclonal antibody MOPC 315 revealed the interaction to have positive ⌬H°and ⌬S°values, and the interaction was thus deduced to be hydrophobically driven (Tanaka et al, 1986). Furthermore, study of the binding of D-glucose to yeast hexokinase showed the enthalpy change (⌬H°) to be nearly zero over the temperature range studied, so that the change in heat capacity was also found to be essentially zero (Takahashi et al, 1981).…”

Characterization of the substitution pattern of cellulose derivatives using carbohydrate-binding modules

“…Affinity gel electrophoresis is a technique that combines the visual simplicity of normal native gel electrophoresis with the specific separation of affinity interactions between various macromolecules. It has been widely used for the binding analysis of various interactions including those between antibody-hapten [ 39 ], lectin-glycoprotein [ 40 ] and between proteins and nucleic acids [ 41 ]. In 1977 AE was used to analyze the binding of a lectin against a soluble polysaccharide [ 42 ] and following the discovery of CBMs this protocol was adopted for studying the interaction of CBMs and polysaccharides [ 43 ].…”