Optimization of oxidized antibody labeling with Lucifer yellow CH

C. R. Keener, C. A.C. Wolfe, D. S. Hage

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17 Scopus citations

Abstract

The oxidation of antibody carbohydrate residues is a common approach used for the site-specific immobilization or modification of antibodies. One way of following this oxidation process is to label the resulting aldehyde groups with a dye such as Lucifer yellow CH (LyCH). This study examined the optimum conditions for preparing and purifying antibody-LyCH conjugates. A 250-fold excess of LyCH reacted with antibody at pH 6.5 for two or more hours gave maximum labeling. Nonreacted LyCH could be effectively removed by passing the labeled antibody through a size exclusion column, followed by one or two dialysis cycles. The LyCH antibody conjugates were found to be stable for at least three weeks when stored in pH 7.4 phosphate buffer.

Original languageEnglish (US)
Pages (from-to)894-895+897
JournalBioTechniques
Volume16
Issue number5
StatePublished - Jan 1 1994

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ASJC Scopus subject areas

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

Cite this

Keener, C. R., Wolfe, C. A. C., & Hage, D. S. (1994). Optimization of oxidized antibody labeling with Lucifer yellow CH. BioTechniques, 16(5), 894-895+897.