Critical evaluation of the applicability of capillary zone electrophoresis for the study of hapten-antibody complex formation.
The applicability of capillary zone electrophoresis (CZE) for the determination of association constants of an hapten-antibody complex with values as high as 10(7) (mol/I)-1 was investigated (dissociation constant in the nmol range). As a reference method the well known, enzyme-linked immunosorbent assay (ELISA) was selected. The study describes the optimisation of the experimental conditions of the CZE technique. The CZE measurements were optimised according to an experimental design. The results of the CZE and ELISA methods are compared giving consideration to the reproducibility (repeatability) of the two methods.
Busch
M H
MH
Laboratory for Analytical Chemistry, Amsterdam Institute for Molecular Studies, University of Amsterdam, The Netherlands.
Boelens
H F
HF
Kraak
J C
JC
Poppe
H
H
Meekel
A A
AA
Resmini
M
M
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000378:metabolism*; D003198:Computer Simulation; D016001:Confidence Intervals; D019075:Electrophoresis, Capillary / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology / Q000378:metabolism*; D008956:Models, Chemical; D009010:Monte Carlo Method; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006241:Haptens