Method to conjugate polysaccharide antigens to surfaces for the detection of antibodies

Ulrik Boas, Peter Lind, Ulla Riber

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    A new generic method for the conjugation of lipopolysaccharide (LPS)-derived polysaccharide antigens from gram-negative bacteria has been developed using Salmonella as a model. After removal of lipid A from the LPS by mild acidolysis, the polysaccharide antigen was conjugated to polystyrene microbeads modified with N-alkyl hydroxylamine and N-alkyl-O-methyl hydroxylamine surface groups by incubation of antigen and beads for 16 h at 40 oC without the need for coupling agents. The efficiency of the new method was evaluated by flow cytometry in model samples and serum samples containing antibodies against Salmonella typhimurium and Salmonella dublin. The presented method was compared with a similar method for conjugation of Salmonella polysaccharide antigens to surfaces. Here, the new method showed higher antigen coupling efficiency by detecting low concentrations of antibodies. Furthermore, the polysaccharide-conjugated beads showed preserved bioactivity after 1 year of use.
    Original languageEnglish
    JournalAnalytical Biochemistry
    Volume465
    Pages (from-to)73-80
    ISSN0003-2697
    DOIs
    Publication statusPublished - 2014

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