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Related Experiment Videos

Recombinant cobra venom factor.

Carl-Wilhelm Vogel1, David C Fritzinger, Brian E Hew

  • 1Cancer Research Center of Hawaii, University of Hawaii, 1236 Lauhala Street, Honolulu, HI 96813, USA. cvogel@crch.hawaii.edu

Molecular Immunology
|May 26, 2004
PubMed
Summary
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Recombinant production of cobra venom factor (CVF) yields active forms, ensuring availability of this crucial complement depletion tool. These methods also enable studies on CVF structure-function and development of humanized CVF for therapies.

Area of Science:

  • Immunology
  • Biochemistry

Background:

  • Cobra venom factor (CVF) is a complement-activating protein functionally similar to C3b but structurally resembling C3c.
  • CVF is vital for depleting complement in animal models to study immune responses and disease pathogenesis.
  • Endangered status of cobras limits natural CVF availability, necessitating alternative production methods.

Purpose of the Study:

  • To review methods for recombinantly producing active forms of cobra venom factor (CVF).
  • To assess the functional activity of recombinant pro-CVF produced in insect cell expression systems.
  • To highlight the importance of recombinant CVF for future research and therapeutic development.

Main Methods:

  • Review of prokaryotic and eukaryotic expression systems for recombinant pro-CVF production.

Related Experiment Videos

  • Utilized baculovirus-infected Sf9 and stably transfected S2 insect cell lines.
  • Characterized three forms of recombinant pro-CVF: single-chain, two-chain (C3-like), and two-chain (C3b-like).

    • Main Results:

    • Insect cell expression systems generated three functional forms of pro-CVF.
    • All recombinant pro-CVF forms exhibited activity indistinguishable from natural CVF.
    • Recombinant pro-CVF formed a C3/C5 convertase (pro-CVF,Bb) that depleted serum complement activity.

    Conclusions:

    • Recombinant production ensures a consistent supply of active CVF for research.
    • Insect cell expression systems are effective for producing functionally active recombinant pro-CVF.
    • Recombinant CVF facilitates studies on complement pathways and the development of humanized CVF for therapeutic applications.