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Single chain antibody (SCA) encoding genes: one-step construction and expression in eukaryotic cells
G T Davis1, W D Bedzyk, E W Voss
1Department of Plant Biology, University of Illinois, Urbana 61801.
Bio/Technology (Nature Publishing Company)
|February 1, 1991
Summary
Researchers developed a fast method to create single chain antibodies (SCAs) in eukaryotic cells. This technique enables the rapid synthesis of functional antigen-binding proteins in various organisms.
Area of Science:
- Molecular Biology
- Immunotechnology
- Biotechnology
Background:
- Single chain antibodies (SCAs) are crucial for various biotechnological applications.
- Efficient gene construction methods are needed for SCA production.
- Expression of functional SCAs in non-lymphatic, non-murine eukaryotic cells remains a challenge.
Purpose of the Study:
- To report the expression of a gene encoding a single chain antibody (SCA) in eukaryotic cells.
- To present a rapid method for constructing genes encoding SCAs.
- To demonstrate the synthesis of functional SCAs in non-lymphatic, non-murine eukaryotic cells.
Main Methods:
- A gene for an SCA targeting fluorescein was constructed using four PCR primers and immunoglobulin cDNA templates.
- Two primers were designed to be partially complementary, encoding the polypeptide linker.
- The constructed gene was expressed in both E. coli and the fission yeast Schizosaccharomyces pombe.
Main Results:
- A functional, hapten-binding SCA product was synthesized in both E. coli and S. pombe.
- The method allows rapid gene synthesis with limited sequence information.
- Murine codon bias, polymerase infidelity, and the eukaryotic environment did not preclude functional SCA synthesis.
Conclusions:
- A rapid and efficient method for constructing SCA genes has been developed.
- Functional SCAs can be synthesized in diverse eukaryotic hosts, including yeast.
- This approach facilitates the production of single chain antigen-binding proteins in non-traditional cellular environments.