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Related Experiment Videos

Fluid-phase SC5b-8 complex of human complement: generation and isolation from serum.

S Bhakdi, M Roth

    Journal of Immunology (Baltimore, Md. : 1950)
    |August 1, 1981
    PubMed
    Summary

    This study details a new method for isolating complement system complexes SC5b-9 and SC5b-8. The technique uses precipitation and chromatography, achieving good recovery and characterizing the SC5b-8 complex.

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    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • The complement system is crucial for innate immunity.
    • Isolation and characterization of complement complexes are vital for understanding their function.

    Purpose of the Study:

    • To develop a rapid and simple method for isolating SC5b-9 and SC5b-8 complement complexes.
    • To characterize the biophysical properties of the isolated SC5b-8 complex.

    Main Methods:

    • Isolation involved polyethyleneglycol precipitation, DEAE ion exchange chromatography, and sucrose density gradient ultracentrifugation.
    • Techniques used for characterization included SDS-gel electrophoresis, gel filtration, and sucrose density gradient centrifugation.

    Main Results:

    • The method achieved approximately 30% recovery for both SC5b-9 and SC5b-8 complexes.

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  • SC5b-8 was identified as a water-soluble macromolecule with alpha-electrophoretic mobility.
  • SC5b-8 exhibited an identical polypeptide pattern to SC5b-9 (minus C9) and a sedimentation coefficient of 19-20S.

    • Conclusions:

    • A robust method for isolating and characterizing complement complexes was established.
    • The SC5b-8 complex was characterized, with a tentative molecular weight of 800-850,000 Da.
    • S-protein remained associated with both complexes, unaffected by desoxycholate treatment.