Design of multivalent complexes using the barnase·barstar module

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

Результат исследований: Материалы для журналаСтатья

143 Цитирования (Scopus)

Выдержка

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.

Язык оригиналаАнглийский
Страницы (с-по)1486-1492
Число страниц7
ЖурналNature Biotechnology
Том21
Номер выпуска12
DOI
СостояниеОпубликовано - дек 2003
Опубликовано для внешнего пользованияДа

Отпечаток

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

Цитировать

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.

В: Nature Biotechnology, Том 21, № 12, 12.2003, стр. 1486-1492.

Результат исследований: Материалы для журналаСтатья

Deyev, SM, Waibel, R, Lebedenko, EN, Schubiger, AP & Plückthun, A 2003, 'Design of multivalent complexes using the barnase·barstar module', Nature Biotechnology, том. 21, № 12, стр. 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 Дек.;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. В: Nature Biotechnology. 2003 ; Том 21, № 12. стр. 1486-1492.
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