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Related Experiment Videos

High-level expression of full-length antibodies using trans-complementing expression vectors.

Allison A Bianchi1, Jeffrey T McGrew

  • 1Amgen Incorporated, 51 University Street, Seattle, WA 98101, USA.

Biotechnology and Bioengineering
|October 24, 2003
PubMed
Summary
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This study introduces a novel mammalian expression system for high-level recombinant antibody production. The system simplifies cell line selection, enabling rapid generation of antibody therapeutics without labor-intensive cloning.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biopharmaceutical Production

Background:

  • Recombinant antibodies are crucial therapeutics, but their production often involves difficult cloning and screening.
  • Existing methods for generating high-expressing cell lines are time-consuming and labor-intensive.

Purpose of the Study:

  • To develop an efficient mammalian expression system for high-level production of full-length antibody molecules.
  • To streamline the generation and selection of cell lines for antibody therapeutics.

Main Methods:

  • Utilized bicistronic vectors linking antibody chain expression to dihydrofolate reductase (DHFR) fragments.
  • Employed a re-associating DHFR selectable marker system with a leucine zipper.
  • Applied methotrexate for concurrent gene amplification and increased antibody production.

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Main Results:

  • Initial pools achieved 5 µg/10⁶ cells/d antibody production.
  • Methotrexate treatment resulted in a two- to fivefold increase in production (10-25 µg/10⁶ cells/d).
  • Shake-flask cultures yielded up to 600 mg/L of antibody within 7 days.

Conclusions:

  • The described system enables rapid antibody generation without cloning, simplifying cell line selection.
  • This approach significantly accelerates the development of potential antibody therapeutics.
  • The method offers a more efficient alternative for biopharmaceutical production.