Recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> produces tailor-made biopolyester granules for applications in fluorescence activated cell sorting: functional display of the mouse interleukin-2 and myelin oligodendrocyte glycoprotein |
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Authors: | B Thomas Bäckström Jane A Brockelbank Bernd HA Rehm |
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Institution: | 1.Malaghan Institute of Medical Research,Wellington,New Zealand;2.Institute of Molecular Biosciences, Massey University,Palmerston North,New Zealand |
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Abstract: | Background Fluorescence activated cell sorting (FACS) is a powerful technique for the qualitative and quantitative detection of biomolecules
used widely in both basic research and clinical diagnostic applications. Beads displaying a specific antigen are used to bind
antibodies which are then fluorescently labelled using secondary antibodies. As the individual suspension bead passes through
the sensing region of the FACS machine, fluorescent signals are acquired and analysed. Currently, antigens are tediously purified
and chemically cross-linked to preformed beads. Purification and coupling of proteins often renders them inactive and they
will not be displayed in its native configuration. As an alternative, we genetically engineered Escherichia coli to produce biopolyester (polyhdroxyalkanoate=PHA) granules displaying diagnostically relevant antigens in their native conformation
and suitable for FACS analysis. |
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