Analysis of human CD4-antibody interaction using the BIAcore system.
Interaction between CD4 cell surface protein and HIV-bearing gp120 has been described as the initial step for HIV entry into host cells. Some anti-CD4 antibodies were shown to inhibit this interaction. Biosensor studies using the BIAcore were performed to determine kinetic and thermodynamic parameters of the interaction of one of these antibodies (i.e. IOT4a, clone 13B8-2) with immobilized recombinant soluble CD4 (rsCD4). A non-linear regression method was used to analyze the sensorgrams, showing the existence of a double exponential time curve. A KA of 5.2 x 10(7) M-1 was calculated at 25 degrees C. The complex formation was exothermic (-4.5 kcal.mol-1( and entropically positive (+20 cal.mol-1.K-1). The reaction rate (0.234 x 10(5) M-1.s-1 at 25 degrees C) as well as the enthalpy change of the activated complex (+9.7 kcal.mol-1) are not compatible with a diffusion controlled reaction. The thermodynamic values calculated from equilibrium data corresponded to those calculated from kinetic data confirming the validity of the theoretical approach. As for most antigen-antibody interactions, complex formation was enthalpy driven. The overall positive entropy contribution to the stabilization of the complex is in contrast to that observed for the lysozyme-anti-lysozyme model and is probably due to electrostatic interaction between the epitope and the antibody combining site.
Velge-Roussel
F
F
CJF INSERM 93-09 Immunologie des Maladies Infectieuses, Equipe Associée INRA d'Immunologie Parasitaire, UFR des Sciences Pharmaceutiques, Tours, France.
Breton
P
P
Lescure
F
F
Guillon
X
X
Bout
D
D
Hoebeke
J
J
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism*; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques; D015704:CD4 Antigens / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D016014:Linear Models; D011485:Protein Binding / Q000502:physiology; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D013816:Thermodynamics
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D011994:Recombinant Proteins