Peak decay analysis and biointeraction studies of immunoglobulin binding and dissociation on protein G affinity microcolumns

Jeanethe A. Anguizola, Erika L. Pfaunmiller, Mitchell L. Milanuk, David S. Hage

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Protein G can be a valuable binding agent for antibodies and immunoglobulins in methods such as immunosensors, chromatographic-based immunoassays, and immunoaffinity chromatography. This report used the method of peak decay analysis along with frontal analysis and zonal elution studies to characterize the binding, elution and regeneration properties of affinity microcolumns that contained immobilized protein G. Frontal analysis was employed with rabbit immunoglobulin G (IgG) to characterize the binding capacity of these affinity microcolumns. Zonal elution experiments looking at the retained peaks for small injections of labeled rabbit IgG were used to optimize the column regeneration conditions. Peak decay analysis was then used to look at the effects of flow rate and elution pH on the release of several types of IgG from the protein G microcolumns. This approach made it possible to obtain detailed information on the use and behavior of such columns, as could be used in future work to optimize the capture or analysis of IgG and antibodies by such devices. The same approach and tools that were used in this report could also be adapted for work with affinity columns that make use of other supports, binding agents or targets.

Original languageEnglish (US)
Pages (from-to)39-45
Number of pages7
JournalMethods
Volume146
DOIs
StatePublished - Aug 15 2018

Keywords

  • Affinity microcolumn
  • Dissociation rate
  • Frontal analysis
  • Immunoglobulin G
  • Peak decay method
  • Protein G

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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