Combinations with Other Methods
The BIACORE system, differential titration calorimetry (DTC) and a stoppedflow instrument have been used for measurement of physico-chemical constants associated with antigen-antibody interactions. After mutations were systematically introduced into the complementarity-determining regions (CDRs) of antibody that correspond to its antigen-binding site, the effects of mutations on its antigen-binding activity were physico-chemically analyzed. Since antigen-antibody interactions are reversible as shown by the following equation,
they must obey the rules of thermodynamics as well as the rules of kinetics. The following three equations show the relationships between rate constants (association rate constant k a, dissociation rate constant k d) and an equilibrium constant (affinity constant K A) as well as the relationships among thermodynamic parameters (∆G, ∆H, ∆S).
$$ Ag + Ab \leftrightarrows Ag \cdot Ab $$
KeywordsNuclear Magnetic Resonance Sedimentation Velocity Nuclear Magnetic Resonance Spectroscopy Sedimentation Coefficient Mass Spectrometry Spectrum
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Unable to display preview. Download preview PDF.
- 6.Glasstone S, Ladler K J, Eyring H (1941) In the Theory of Rate Processes. McGraw-Hill, New YorkGoogle Scholar
- 9.Busch K, Glish G L, McLuckey S A (1988) Mass Spectrometry/ Mass Spectrometry: Techniques and Applications of Tandem Mass Spectrometry. VCH, New YorkGoogle Scholar
- 11.Weber-Grabauy M, Kelley P, Syka J, Bradshaw S, Brodbelt J (1987) Proc. 35th ASMS Conf. Mass Spectrom. Allied Topics, Denver, COGoogle Scholar
- 12.March R E, Hughes R J (1989) Quadrupole Storage Mass Spectrometry. Wiley, New YorkGoogle Scholar
- 15.Lackmann M, Bucci T, Mann R J, Kravets L A, Viney E, Smith F, Moritz R L, Carter W, Simpson R J, Nicola N A, Mackwell K, Nice E C, Wilks A F, Boyd A W (1996) Purification of the EPH-like receptor HEK using a biosensor-based affinity detection approach. Proc. Natl Acad. Sci. USA 93: 2523–2527PubMedCrossRefGoogle Scholar
- 1.Wüthrich K (1986) NMR of Proteins and Nucleic Acids. John Wiley & SonsGoogle Scholar
- 2.Kainosho M, Tsuji T ( 1982) Assignment of the three methionyl carbonyl carbon resonances in Streptomyces subtilisin inhibitor by a carbon-13 and nitrogen-15 double-labeling technique. A new strategy for structural studies of proteins in solution. Biochemistry 21: 6273–6279PubMedCrossRefGoogle Scholar
- 5.Nakayama T, Arata A, Shimada I (1993) A Multinuclear NMR Study of the Affinity Maturation of Anti-NP Mouse Monoclonal Antibodies: Comparison of Antibody Combining Sites between Primary Response Antibody N1G9 and Secondary Response Antibody 3B62. Biochemistry 32: 13961–13968PubMedCrossRefGoogle Scholar
- 2.Ralston G (1993) Introduction to Analytical Ultracentrifugation. Beckman Instruments, FullertonGoogle Scholar
- 3.Laue T, Philo J, Hays D. SEDNTERP: available on rasmb ftp site.Google Scholar
- 7.Cantor C R, Schimmel P R (1980) Biophysical Chemistry Part II. Techniques for the study of biological structure and function, pp.555–570, 610–612Google Scholar
- 10.McRorie D K, Voelker P J (1993) Self-Associating Systems in the Analytical Ultracentri-fuge. Beckman Instruments, FullertonGoogle Scholar
© Springer Japan 2000