A synthetic trivalent hapten that aggregates anti-2,4-DNP IgG into bicyclic trimers.
This paper describes the synthesis of the trivalent hapten molecule 1, containing three 2,4-dinitrophenyl (2,4-DNP) groups, and the use of this molecule to aggregate three molecules of anti-2,4-DNP IgG into a complex with 3:2 stoichiometry (IgG312). The equilibrium product IgG312 was generated in approximately 90% yield upon mixing IgG and 1; during incubation, thermodynamically unstable, high-molecular-weight aggregates (>104 nm in diameter) form first and convert subsequently to IgG312. The thermodynamics and the kinetics of the formation of aggregates were studied using size-exclusion high-performance liquid chromatography (SE-HPLC), dynamic light scattering (DLS), and analytical ultracentrifugation (AUC). An analytical model based on multiple species in equilibrium was developed and used to interpret the SE-HPLC data. The aggregate IgG312 was more stable thermodynamically and kinetically than monomeric aggregates of this IgG with monomeric derivatives of 2,4-DNP; this stability suggests potential applications of these aggregates in biotechnology.
Bilgiçer
Basar
B
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.
Moustakas
Demetri T
DT
Whitesides
George M
GM
D019297:2,4-Dinitrophenol / Q000276:immunology*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006241:Haptens / Q000737:chemistry* / Q000276:immunology*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology*; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D051381:Rats; D013816:Thermodynamics; D014461:Ultracentrifugation
D006241:Haptens; D007074:Immunoglobulin G; D019297:2,4-Dinitrophenol