Application of surface plasmon resonance toward studies of low-molecular-weight antigen-antibody binding interactions
被引:74
作者:
Adamczyk, M
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h-index: 0
机构:
Abbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USAAbbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USA
Adamczyk, M
[1
]
Moore, JA
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h-index: 0
机构:
Abbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USAAbbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USA
Moore, JA
[1
]
Yu, ZG
论文数: 0引用数: 0
h-index: 0
机构:
Abbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USAAbbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USA
Yu, ZG
[1
]
机构:
[1] Abbott Labs, Diagnost Div Organ Chem 9NM, Abbott Pk, IL 60064 USA
来源:
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
|
2000年
/
20卷
/
03期
关键词:
D O I:
10.1006/meth.1999.0925
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
Methods for studying low-molecular-weight antigen-antibody binding interactions using surface plasmon resonance detection are presented. The experimental parameters most relevant to studies of low-molecular-weight antigen-antibody binding interactions are discussed. Direct kinetic analysis of the binding interactions is most informative, providing both apparent association and dissociation rate constants from which equilibrium constants can be calculated. Equilibrium analysis, including steady-state and solution affinity studies, offers an alternative approach to direct kinetic analysis when knowledge of the individual kinetic rate constants is not required or difficult to determine. The various methods are illustrated by studies of an anti-T-4 Fab fragment binding interaction with several thyroxine analogs. The methods utilized were dependent on the affinity of the interaction. The high-affinity anti-T-4 Fab fragment/L-T-4 binding interaction was evaluated using direct kinetic analysis. An intermediate affinity anti-T-4 Fab fragment/L-T-3 binding interaction was evaluated using a combination of direct kinetic analysis, steady-state analysis, and solution affinity analysis. The relatively weak anti-T-4 Fab fragment/L-T-3 binding interaction was evaluated using steady-state and solution affinity analysis protocols. Several thyroxine tracers that could not be immobilized to a biosensor surface were also evaluated via the solution affinity format. In cases where a given binding interaction was examined using multiple methods the results were comparable. (C) 2000 Academic Press.
机构:
GENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USAGENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USA
CUNNINGHAM, BC
WELLS, JA
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机构:
GENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USAGENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USA
机构:
GENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USAGENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USA
CUNNINGHAM, BC
WELLS, JA
论文数: 0引用数: 0
h-index: 0
机构:
GENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USAGENENTECH INC, DEPT PROT ENGN, 460 POINT SAN BRUNO BLVD, S SAN FRANCISCO, CA 94080 USA