High-throughput immunoturbidimetric assays for in-process determination of polyclonal antibody concentration and functionality in crude samples

Hanne Bak, J. Kyhse-Andersen, O.R.T. Thomas

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    We present fast, simple immunoturbidimetric assays suitable for direct determination of antibody 'concentration' and 'functionality' in crude samples, such as in-process samples taken at various stages during antibody purification. Both assays display excellent linearity and analytical recovery. Possible influences from commonly employed buffers and salts (present in samples at various concentrations), and of pH variations, were studied for both assays. Interference effects were shown to be negligible for the 'concentration' assay, such that sample pre-treatment prior to assay is unnecessary. The 'functionality' assay displayed concentration dependent sensitivity to interference for ammonium sulphate and Tris(hydroxymethyl)-amino-methane, but was essentially unaffected by all other salts and buffer combinations tested. The immunoturbidimetric assays described here are generically applicable to polyclonal antibodies, require only basic laboratory equipment, are robust, fast, cheap, easy to perform, and readily adapted to automation.
    Original languageEnglish
    JournalJournal of Chromatography B-analytical Technologies in the Biomedical and Life Sciences
    Volume848
    Issue number1
    Pages (from-to)142-150
    ISSN1570-0232
    DOIs
    Publication statusPublished - 2007

    Keywords

    • immunoglobulins
    • precipitin reaction
    • turbidimetry
    • nephelometry
    • titre

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