Use of uteroglobin for the engineering of polyvalent, polyspecific fusion proteins

Autor: Sara Fossati, Barbara Carnemolla, Arianna Parodi, Enrica Balza, William L. Blalock, Luciano Zardi, Francesca Sassi, Elisa Ventura, Laura Borsi, Patrizia Castellani
Rok vydání: 2009
Předmět:
Cytotoxicity
Immunologic
Models
Molecular
Secondary
Cytotoxicity
Inbred Strains
Immunoglobulin Variable Region
Biochemistry
Protein Structure
Secondary
law.invention
Mice
Immunologic
Models
law
Antibody Specificity
Neoplasms
Monoclonal
Uteroglobin
Tumor
biology
Proteolytic enzymes
Antibodies
Monoclonal
Biological activity
Protein Structure and Folding
Recombinant DNA
Electrophoresis
Polyacrylamide Gel
Antibody
Oxidation-Reduction
Plasmids
Electrophoresis
Protein Structure
Recombinant Fusion Proteins
Enzyme-Linked Immunosorbent Assay
Mice
Inbred Strains
Protein Sorting Signals
Antibodies
Cell Line
Experimental
Cell Line
Tumor
Animals
Humans
Molecular Biology
Polyacrylamide Gel
Tumor Necrosis Factor-alpha
Molecular
Cell Biology
Neoplasms
Experimental
Fusion protein
Molecular biology
Fibronectin
biology.protein
Interleukin-2
Protein Multimerization
Zdroj: The Journal of biological chemistry. 284(39)
ISSN: 1083-351X
Popis: We report a novel strategy to engineer and express stable and soluble human recombinant polyvalent/polyspecific fusion proteins. The procedure is based on the use of a central skeleton of uteroglobin, a small and very soluble covalently linked homodimeric protein that is very resistant to proteolytic enzymes and to pH variations. Using a human recombinant antibody (scFv) specific for the angiogenesis marker domain B of fibronectin, interleukin 2, and an scFv able to neutralize tumor necrosis factor-alpha, we expressed various biologically active uteroglobin fusion proteins. The results demonstrate the possibility to generate monospecific divalent and tetravalent antibodies, immunocytokines, and dual specificity tetravalent antibodies. Furthermore, compared with similar fusion proteins in which uteroglobin was not used, the use of uteroglobin improved properties of solubility and stability. Indeed, in the reported cases it was possible to vacuum dry and reconstitute the proteins without any aggregation or loss in protein and biological activity.
Databáze: OpenAIRE
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