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  • ZZ polyester beads: An efficient and simple method for purifying IgG from mouse hybridoma supernatants

    Author(s)
    Lewis, John G
    Rehm, Bernd HA
    Griffith University Author(s)
    Rehm, Bernd
    Year published
    2009
    Metadata
    Show full item record
    Abstract
    ZZ polyester beads are polyhydroxyalkanoate granules which display, at high density, the ZZ domain of protein A as a fusion protein covalently linked to the polyester core. These granules are produced directly, in one step, by recombinant E. coli. We have used these granules produced in a one step process to purify mouse IgG1, IgG2a and IgG2b from mouse hybridoma supernatants. Purified immunoglobulins were analysed by enzyme-linked immonosorbent assay, agarose gel electrophoresis and SDS-PAGE. The results showed that the recovery of IgG is 70% or greater with a significant degree of purity. ZZ polyester beads hence offer a ...
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    ZZ polyester beads are polyhydroxyalkanoate granules which display, at high density, the ZZ domain of protein A as a fusion protein covalently linked to the polyester core. These granules are produced directly, in one step, by recombinant E. coli. We have used these granules produced in a one step process to purify mouse IgG1, IgG2a and IgG2b from mouse hybridoma supernatants. Purified immunoglobulins were analysed by enzyme-linked immonosorbent assay, agarose gel electrophoresis and SDS-PAGE. The results showed that the recovery of IgG is 70% or greater with a significant degree of purity. ZZ polyester beads hence offer a rapid and novel method to purify IgG from mouse hybridoma culture supernatants. As a negative control we used culture supernatant from an IgM secreting hybridoma and showed that it did not bind to ZZ polyester beads.
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    Journal Title
    Journal of Immunological Methods
    Volume
    346
    Issue
    1-2
    DOI
    https://doi.org/10.1016/j.jim.2009.04.011
    Subject
    Immunology
    Immunology not elsewhere classified
    Medical microbiology
    Ophthalmology and optometry
    Publication URI
    http://hdl.handle.net/10072/350785
    Collection
    • Journal articles

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