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  Reference Spotlight

In a 2012 publication from PLoS One, researchers investigated the dynamic range of the Kinetic Exclusion Assay (KinExA) for the characterization of antigen-antibody interactions. Due to the versatility of KinExA, experiments could be run using recombinant or unpurified native antigens. By using the standard and reverse assay formats, the amount of material required for the experiments could be minimized. Anti-idiotypes were used when applicable to help reduce material usage and cost. The range of Kd values for the tested experimental systems spanned six orders of magnitude, from approximately 100 fM to 100 nM.

Bee Christine, et al. 2012. Exploring the dynamic range of the kinetic exclusion assay in characterizing antigen-antibody interactions. PLOS ONE 7(4): e36261 http://www.ncbi.nlm.nih.gov/pubmed/22558410

KinExA References

Affinity & Kinetic Measurements:

KinExA technology overview:

KinExA’s role in drug discovery:

Significance of “solution phase” measurements to unmodified molecules:

Sensitivity to measure tight binders:

Reverse assay techniques:

Whole cell binding techniques:

  • Bedinger, D., et al. 2015. Differential pathway coupling of activated insulin receptor drives signaling selectivity by XmetA, an allosteric partial agonist antibody. J Pharmacol Exp Ther 353(1):35-43. http://www.ncbi.nlm.nih.gov/pubmed/25613982
  • Rathanaswami P., Babcook J., Gallo M. 2008. High-affinity binding measurements of antibodies to cell-surface-expressed antigens. Anal Biochem 373: 52-60. http://www.ncbi.nlm.nih.gov/pubmed/17910940
  • Xie L., et al. 2005. Measurement of the functional affinity constant of a monoclonal antibody for cell surface receptors using kinetic exclusion fluorescence immunoassay. J Immunol Methods 304: 1-14. http://www.ncbi.nlm.nih.gov/pubmed/16098983

Unpurified antigens:

Other interesting studies:


Immunoassay Techniques:

  • Darwish I.A., et al. 2013. Kinetic-exclusion analysis-based immunosensors versus enzyme-linked immunosorbent assays for measurement of cancer markers in biological specimens. Talanta 111: 13-19. http://www.ncbi.nlm.nih.gov/pubmed/23622520
  • Prieto-Simon B., Miyachi H., Karube I., Saiki H. 2010. High-sensitive flow-based kinetic exclusion assay for okadaic acid assessment in shellfish samples. Biosens Bioelectron 25: 1395-1401. http://www.ncbi.nlm.nih.gov/pubmed/19939663
  • Sasaki K., Oguma S., Namiki Y., Ohmura N. 2009. Monoclonal antibody to trivalent chromium chelate complex and its application to measurement of the total chromium concentration. Anal Chem 81: 4005-4009. http://www.ncbi.nlm.nih.gov/pubmed/19438265
  • Glass T.R., Ohmura N., Saiki H. 2007. Least detectable concentration and dynamic range of three immunoassay systems using the same antibody. Anal Chem 79: 1954-1960. http://www.ncbi.nlm.nih.gov/pubmed/17256970
  • Bromage E.S., et al. 2007. The development of a real-time biosensor for the detection of trace levels of trinitrotoluene (TNT) in aquatic environments. Biosens Bioelectron 22: 2532-2538. http://www.ncbi.nlm.nih.gov/pubmed/17088054
  • Sasaki K., Glass T.R., Ohmura N. 2005. Validation of accuracy of enzyme-linked immunosorbent assay in hybridoma screening and proposal of an improved screening method. Anal Chem 77: 1933-1939. http://www.ncbi.nlm.nih.gov/pubmed/15801721
  • Glass T.R., et al. 2004. Use of excess solid-phase capacity in immunoassays: advantages for semicontinuous, near-real-time measurements and for analysis of matrix effects. Anal Chem 76: 767-772. http://www.ncbi.nlm.nih.gov/pubmed/14750874
  • Ohmura N., Lackie S., Saiki H. 2001. An immunoassay for small analytes with theoretical detection limits. Anal Chem 73: 3392-3399. http://www.ncbi.nlm.nih.gov/pubmed/11476240