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. 2000 Dec;79(6):2954–2965. doi: 10.1016/S0006-3495(00)76532-5

Kinetics of association of anti-lysozyme monoclonal antibody D44.1 and hen-egg lysozyme.

G Altobelli 1, S Subramaniam 1
PMCID: PMC1301174  PMID: 11106603

Abstract

Association rate constants for antigen/antibody associations have been computed by Brownian Dynamics simulations of D. L. Ermak and J. A. McCammon, J. Chem. Phys. 69:1352-1360, 1978. The model of monoclonal antibody (mAb) D44.1 is based on crystallographic data (B. C. Braden et al., J. Mol. Biol. 243:767-781, 1994). Electrostatic forces that steer the antigen to the antibody-combining site are computed by solving the linearized Poisson-Boltzmann equation. D44. 1-HEL complex displays very similar association motifs to a related anti-lysozyme antibody, HyHEL-5-HEL system. The computed association rate constants are comparable in the two systems, although the experimental affinity constants differ by three orders of magnitude (D. Tello et al., Biochem. Soc. Trans. 21:943-946, 1993; K. A. Hibbits et al., Biochemistry. 33:3584-3590, 1994). Simulations suggest that the origin of the differences in the affinity come from dissociation rate constants. We have also carried out simulation experiments on a number of mutant antibody fragment-HEL associations to address the role of electrostatics and, to a limited extent, the orientational aspects of association.

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Selected References

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  1. Batista F. D., Neuberger M. S. Affinity dependence of the B cell response to antigen: a threshold, a ceiling, and the importance of off-rate. Immunity. 1998 Jun;8(6):751–759. doi: 10.1016/s1074-7613(00)80580-4. [DOI] [PubMed] [Google Scholar]
  2. Berg O. G., von Hippel P. H. Diffusion-controlled macromolecular interactions. Annu Rev Biophys Biophys Chem. 1985;14:131–160. doi: 10.1146/annurev.bb.14.060185.001023. [DOI] [PubMed] [Google Scholar]
  3. Bhat T. N., Bentley G. A., Fischmann T. O., Boulot G., Poljak R. J. Small rearrangements in structures of Fv and Fab fragments of antibody D1.3 on antigen binding. Nature. 1990 Oct 4;347(6292):483–485. doi: 10.1038/347483a0. [DOI] [PubMed] [Google Scholar]
  4. Braden B. C., Poljak R. J. Structural features of the reactions between antibodies and protein antigens. FASEB J. 1995 Jan;9(1):9–16. doi: 10.1096/fasebj.9.1.7821765. [DOI] [PubMed] [Google Scholar]
  5. Braden B. C., Souchon H., Eiselé J. L., Bentley G. A., Bhat T. N., Navaza J., Poljak R. J. Three-dimensional structures of the free and the antigen-complexed Fab from monoclonal anti-lysozyme antibody D44.1. J Mol Biol. 1994 Nov 4;243(4):767–781. doi: 10.1016/0022-2836(94)90046-9. [DOI] [PubMed] [Google Scholar]
  6. Davies D. R., Cohen G. H. Interactions of protein antigens with antibodies. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):7–12. doi: 10.1073/pnas.93.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. EISEN H. N., SISKIND G. W. VARIATIONS IN AFFINITIES OF ANTIBODIES DURING THE IMMUNE RESPONSE. Biochemistry. 1964 Jul;3:996–1008. doi: 10.1021/bi00895a027. [DOI] [PubMed] [Google Scholar]
  8. Eisen H. N. The immune response to a simple antigenic determinant. Harvey Lect. 1966;60:1–34. [PubMed] [Google Scholar]
  9. Foote J., Eisen H. N. Kinetic and affinity limits on antibodies produced during immune responses. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1254–1256. doi: 10.1073/pnas.92.5.1254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Foote J., Milstein C. Kinetic maturation of an immune response. Nature. 1991 Aug 8;352(6335):530–532. doi: 10.1038/352530a0. [DOI] [PubMed] [Google Scholar]
  11. Gabdoulline R. R., Wade R. C. On the protein-protein diffusional encounter complex. J Mol Recognit. 1999 Jul-Aug;12(4):226–234. doi: 10.1002/(SICI)1099-1352(199907/08)12:4<226::AID-JMR462>3.0.CO;2-P. [DOI] [PubMed] [Google Scholar]
  12. Gabdoulline R. R., Wade R. C. Simulation of the diffusional association of barnase and barstar. Biophys J. 1997 May;72(5):1917–1929. doi: 10.1016/S0006-3495(97)78838-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibas C. J., Subramaniam S., McCammon J. A., Braden B. C., Poljak R. J. pH dependence of antibody/lysozyme complexation. Biochemistry. 1997 Dec 16;36(50):15599–15614. doi: 10.1021/bi9701989. [DOI] [PubMed] [Google Scholar]
  14. Hibbits K. A., Gill D. S., Willson R. C. Isothermal titration calorimetric study of the association of hen egg lysozyme and the anti-lysozyme antibody HyHEL-5. Biochemistry. 1994 Mar 29;33(12):3584–3590. doi: 10.1021/bi00178a015. [DOI] [PubMed] [Google Scholar]
  15. Kozack R. E., Subramaniam S. Brownian dynamics simulations of molecular recognition in an antibody-antigen system. Protein Sci. 1993 Jun;2(6):915–926. doi: 10.1002/pro.5560020605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kozack R. E., d'Mello M. J., Subramaniam S. Computer modeling of electrostatic steering and orientational effects in antibody-antigen association. Biophys J. 1995 Mar;68(3):807–814. doi: 10.1016/S0006-3495(95)80257-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  18. Newman M. A., Mainhart C. R., Mallett C. P., Lavoie T. B., Smith-Gill S. J. Patterns of antibody specificity during the BALB/c immune response to hen eggwhite lysozyme. J Immunol. 1992 Nov 15;149(10):3260–3272. [PubMed] [Google Scholar]
  19. Nicholls A., Sharp K. A., Honig B. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins. 1991;11(4):281–296. doi: 10.1002/prot.340110407. [DOI] [PubMed] [Google Scholar]
  20. Northrup S. H., Erickson H. P. Kinetics of protein-protein association explained by Brownian dynamics computer simulation. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3338–3342. doi: 10.1073/pnas.89.8.3338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Raman C. S., Jemmerson R., Nall B. T., Allen M. J. Diffusion-limited rates for monoclonal antibody binding to cytochrome c. Biochemistry. 1992 Oct 27;31(42):10370–10379. doi: 10.1021/bi00157a027. [DOI] [PubMed] [Google Scholar]
  22. Roost H. P., Bachmann M. F., Haag A., Kalinke U., Pliska V., Hengartner H., Zinkernagel R. M. Early high-affinity neutralizing anti-viral IgG responses without further overall improvements of affinity. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1257–1261. doi: 10.1073/pnas.92.5.1257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schwarz F. P., Tello D., Goldbaum F. A., Mariuzza R. A., Poljak R. J. Thermodynamics of antigen-antibody binding using specific anti-lysozyme antibodies. Eur J Biochem. 1995 Mar 1;228(2):388–394. [PubMed] [Google Scholar]
  24. Tello D., Goldbaum F. A., Mariuzza R. A., Ysern X., Schwarz F. P., Poljak R. J. Three-dimensional structure and thermodynamics of antigen binding by anti-lysozyme antibodies. Biochem Soc Trans. 1993 Nov;21(4):943–946. doi: 10.1042/bst0210943. [DOI] [PubMed] [Google Scholar]
  25. Wade R. C. Brownian dynamics simulations of enzyme-substrate encounter. Biochem Soc Trans. 1996 Feb;24(1):254–259. doi: 10.1042/bst0240254. [DOI] [PubMed] [Google Scholar]
  26. Warwicker J., Watson H. C. Calculation of the electric potential in the active site cleft due to alpha-helix dipoles. J Mol Biol. 1982 Jun 5;157(4):671–679. doi: 10.1016/0022-2836(82)90505-8. [DOI] [PubMed] [Google Scholar]
  27. Winter G., Milstein C. Man-made antibodies. Nature. 1991 Jan 24;349(6307):293–299. doi: 10.1038/349293a0. [DOI] [PubMed] [Google Scholar]
  28. Xavier K. A., Willson R. C. Association and dissociation kinetics of anti-hen egg lysozyme monoclonal antibodies HyHEL-5 and HyHEL-10. Biophys J. 1998 Apr;74(4):2036–2045. doi: 10.1016/s0006-3495(98)77910-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zhou H. X. Brownian dynamics study of the influences of electrostatic interaction and diffusion on protein-protein association kinetics. Biophys J. 1993 Jun;64(6):1711–1726. doi: 10.1016/S0006-3495(93)81543-1. [DOI] [PMC free article] [PubMed] [Google Scholar]

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