Confirmation of the validity of the current characterization of immunochemical reactions by kinetic exclusion assay

Glass, Thomas R. and Winzor, Donald J. (2014) Confirmation of the validity of the current characterization of immunochemical reactions by kinetic exclusion assay. Analytical Biochemistry, 456 1: 38-42. doi:10.1016/j.ab.2014.04.011


Author Glass, Thomas R.
Winzor, Donald J.
Title Confirmation of the validity of the current characterization of immunochemical reactions by kinetic exclusion assay
Journal name Analytical Biochemistry   Check publisher's open access policy
ISSN 0003-2697
1096-0309
Publication date 2014-07-01
Sub-type Article (original research)
DOI 10.1016/j.ab.2014.04.011
Open Access Status
Volume 456
Issue 1
Start page 38
End page 42
Total pages 5
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Collection year 2015
Language eng
Formatted abstract
Prior observations that questioned the validity of kinetic exclusion assays were based on the mistaken assumption that the assays quantified the fraction of those antibody molecules that had unoccupied binding sites. Instead, the standard KinExA assay quantifies the fraction of total antibody binding sites that are unoccupied, regardless of the number of unoccupied sites on each antibody molecule. Although the standard KinExA analysis assumes that there is only a small probability of antibody-site capture by the affinity matrix, the results of numerical simulations demonstrate the reliability of dissociation constants obtained by the standard KinExA analysis for capture probabilities as high as 30%. This finding further strengthens the potential of kinetic exclusion assays as the procedure of choice for the rapid and accurate characterization of immunochemical reactions that forms part of screening processes in the search for therapeutic antibodies.
Keyword Antigen–antibody interactions
Antibody bivalence
Kinetic exclusion assay
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Chemistry and Molecular Biosciences
 
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Created: Mon, 05 May 2014, 15:32:30 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences