Design of multivalent complexes using the barnase·barstar module

Sergey M. Deyev, Robert Waibel, Ekaterina N. Lebedenko, August P. Schubiger, Andreas Plückthun

Research output: Contribution to journalArticle

141 Citations (Scopus)

Abstract

The ribonuclease barnase (12 kDa) and its inhibitor barstar (10 kDa) form a very tight complex in which all N and C termini are accessible for fusion. Here we exploit this system to create modular targeting molecules based on antibody scFv fragment fusions to barnase, to two barnase molecules in series and to barstar. We describe the construction, production and purification of defined dimeric and trimeric complexes. Immobilized barnase fusions are used to capture barstar fusions from crude extracts to yield homogeneous, heterodimeric fusion proteins. These proteins are stable, soluble and resistant to proteolysis. Using fusions with anti-p185HER2-ECD 4D5 scFv, we show that the anticipated gain in avidity from monomer to dimer to trimer is obtained and that favorable tumor targeting properties are achieved. Many permutations of engineered multispecific fusion proteins become accessible with this technology of quasi-covalent heterodimers.

Original languageEnglish
Pages (from-to)1486-1492
Number of pages7
JournalNature Biotechnology
Volume21
Issue number12
DOIs
Publication statusPublished - Dec 2003
Externally publishedYes

Fingerprint

Fusion reactions
Immunoglobulin Fragments
Proteins
Ribonucleases
Complex Mixtures
Proteolysis
Single-Chain Antibodies
Molecules
Technology
Bacillus amyloliquefaciens ribonuclease
Antibodies
Dimers
Purification
Tumors
Bacillus amyloliquefaciens barstar protein
Monomers
Neoplasms

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering
  • Molecular Medicine

Cite this

Deyev, S. M., Waibel, R., Lebedenko, E. N., Schubiger, A. P., & Plückthun, A. (2003). Design of multivalent complexes using the barnase·barstar module. Nature Biotechnology, 21(12), 1486-1492. https://doi.org/10.1038/nbt916

Design of multivalent complexes using the barnase·barstar module. / Deyev, Sergey M.; Waibel, Robert; Lebedenko, Ekaterina N.; Schubiger, August P.; Plückthun, Andreas.

In: Nature Biotechnology, Vol. 21, No. 12, 12.2003, p. 1486-1492.

Research output: Contribution to journalArticle

Deyev, SM, Waibel, R, Lebedenko, EN, Schubiger, AP & Plückthun, A 2003, 'Design of multivalent complexes using the barnase·barstar module', Nature Biotechnology, vol. 21, no. 12, pp. 1486-1492. https://doi.org/10.1038/nbt916
Deyev SM, Waibel R, Lebedenko EN, Schubiger AP, Plückthun A. Design of multivalent complexes using the barnase·barstar module. Nature Biotechnology. 2003 Dec;21(12):1486-1492. https://doi.org/10.1038/nbt916
Deyev, Sergey M. ; Waibel, Robert ; Lebedenko, Ekaterina N. ; Schubiger, August P. ; Plückthun, Andreas. / Design of multivalent complexes using the barnase·barstar module. In: Nature Biotechnology. 2003 ; Vol. 21, No. 12. pp. 1486-1492.
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