Studies on the rate and control of antibody oxidation by periodate

Carrie A.C. Wolfe, David S. Hage

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

The oxidation of antibody carbohydrate residues by periodate is a common approach for the site-specific immobilization or modification of antibodies for use in various bioanalytical methods. This study examined the time dependence of this oxidation process under a variety of pH, temperature, and concentration conditions. Polyclonal rabbit immunoglobulin G (IgG) was used as the model system for these studies. Flow-injection analysis and a hydrazide label (Lucifer yellow CH) were used to monitor the progress of the oxidation reaction. It was found that the number of oxidized sites that were available for labeling could be varied between one and eight groups per antibody by adjusting the time, pH, periodate concentration, or reaction temperature. In each case, most of these groups were produced during the first 30-60 min of the reaction. A comparison was made between these results and those of previous studies that have examined the effects of periodate treatment on amino acid residues and antibody activity. From this work, general guidelines were developed for the control and optimization of antibody oxidation for use with assays that require either high or low levels of antibody modification.

Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalAnalytical Biochemistry
Volume231
Issue number1
DOIs
StatePublished - Oct 1995

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Oxidation
Antibodies
Flow Injection Analysis
Temperature
Immobilization
Labeling
metaperiodate
Labels
Assays
Immunoglobulin G
Carbohydrates
Guidelines
Rabbits
Amino Acids

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Studies on the rate and control of antibody oxidation by periodate. / Wolfe, Carrie A.C.; Hage, David S.

In: Analytical Biochemistry, Vol. 231, No. 1, 10.1995, p. 123-130.

Research output: Contribution to journalArticle

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