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An efficient method for introducing macromolecules into living cells.

S J Doxsey, J Sambrook, A Helenius

    The Journal of Cell Biology
    |July 1, 1985
    PubMed
    Summary
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    Influenza hemagglutinin (HA) enables efficient delivery of antibodies and HRP into cells using protein-loaded erythrocytes. This novel, nonlytic method ensures high delivery rates without compromising cell viability for biochemical analysis.

    Area of Science:

    • Cell Biology
    • Virology
    • Biotechnology

    Background:

    • Influenza virus hemagglutinin (HA) mediates cell entry through membrane fusion.
    • Efficient delivery of macromolecules into living cells remains a challenge in biotechnology.

    Purpose of the Study:

    • To develop a novel method for rapid, bulk delivery of antibodies and horseradish peroxidase (HRP) into the cytoplasm of living cells.
    • To utilize influenza hemagglutinin's properties for targeted cell entry and fusion.

    Main Methods:

    • Utilized cells engineered to express influenza hemagglutinin (HA) on their surface.
    • Bound protein-loaded erythrocytes to HA-expressing cells.
    • Triggered erythrocyte-cell fusion using an acid-dependent pH drop (pH 5.0).

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    Main Results:

    • Achieved efficient delivery of IgG and HRP into 75-95% of recipient cells.
    • Delivered large quantities of HRP (up to 1.8 x 10^8 molecules/cell) and IgG (1.4 x 10^7 molecules/cell).
    • Confirmed no impairment of cell viability, growth, or division post-delivery.

    Conclusions:

    • The HA-mediated erythrocyte fusion method provides simple, reliable, and nonlytic delivery of impermeable substances into cells.
    • This technique facilitates rapid, simultaneous delivery into large cell populations for biochemical studies.
    • Fused erythrocyte membranes were observed as discrete domains within the plasma membrane, indicating successful integration.