Radiometal labeling of recombinant proteins by a genetically engineered minimal chelation site: technetium-99m coordination by single-chain Fv antibody fusion proteins through a C-terminal cysteinyl peptide
| Autor: | F Jamar, M S Tai, Louis M. Weiner, L L Houston, A. M. Peters, Hermann Oppermann, G P Adams, B T Heelan, Andrew J.T. George, John E. McCartney |
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| Rok vydání: | 1995 |
| Předmět: |
Male
Protein Folding Recombinant Fusion Proteins viruses medicine.medical_treatment Molecular Sequence Data Peptide Mice SCID law.invention Mice Antigen law medicine Animals Tissue Distribution Amino Acid Sequence Cysteine Cloning Molecular Radionuclide Imaging Immunoglobulin Fragments Peptide sequence Chelating Agents chemistry.chemical_classification Multidisciplinary biology C-terminus Technetium virus diseases Neoplasms Experimental biochemical phenomena metabolism and nutrition Molecular biology Fusion protein Rats Kinetics chemistry Biochemistry Radioimmunotherapy biology.protein Recombinant DNA Antibody Peptides Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences. 92:8358-8362 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.92.18.8358 |
| Popis: | We describe a method to facilitate radioimaging with technetium-99m (99mTc) by genetic incorporation of a 99mTc chelation site in recombinant single-chain Fv (sFv) antibody proteins. This method relies on fusion of the sFv C terminus with a Gly4Cys peptide that specifically coordinates 99mTc. By using analogues of the 26-10 anti-digoxin sFv as our primary model, we find that addition of the chelate peptide, to form 26-10-1 sFv', does not alter the antigen-binding affinity of sFv. We have demonstrated nearly quantitative chelation of 0.5-50 mCi of 99mTc per mg of 26-10-1 sFv' (1 Ci = 37 GBq). These 99mTc-labeled sFv' complexes are highly stable to challenge with saline buffers, plasma, or diethylenetriaminepentaacetic acid. We find that the 99mTc-labeled 741F8-1 sFv', specific for the c-erbB-2 tumor-associated antigen, is effective in imaging human ovarian carcinoma in a scid mouse tumor xenograft model. This fusion chelate methodology should be applicable to diagnostic imaging with 99mTc and radioimmunotherapy with 186Re or 188Re, and its use could extend beyond the sFv' to other engineered antibodies, recombinant proteins, and synthetic peptides. |
| Databáze: | OpenAIRE |
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