Antigenicity of recombinant proteins after regioselective immobilization onto polyanhydride-based copolymers
| Autor: | François Mallet, Emmanuelle Imbert-Laurenceau, Laure Allard, Bernard Mandrand, Guy Oriol, Thierry Delair, Valérie Cheynet, Gaspard Gervasi |
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| Rok vydání: | 2004 |
| Předmět: |
Antigenicity
Polymers Lysine Molecular Sequence Data Biomedical Engineering HIV Core Protein p24 Pharmaceutical Science Bioengineering Peptide Mapping Anhydrides Hydrolysis Epitopes medicine Copolymer Organic chemistry Amino Acid Sequence Pharmacology Bioconjugation Chemistry Organic Chemistry Stereoisomerism Vinyl ether Combinatorial chemistry Recombinant Proteins Immobilized Proteins Covalent bond Amine gas treating Biotechnology medicine.drug |
| Zdroj: | Bioconjugate chemistry. 15(3) |
| ISSN: | 1043-1802 |
| Popis: | We previously demonstrated that the introduction of a tag consisting of several contiguous lysines at the N- or C-terminus of a recombinant protein greatly improved the covalent grafting of the protein onto negatively charged maleic anhydride-alt-methyl vinyl ether (MAMVE) copolymer, under many different experimental conditions (Ladaviere, C., et al. (1998) Bioconjugate Chem. 9, 655; Allard, L., et al. (2002) Biotechnol. Bioeng. 80, 341). The grafting efficiency was dependent on the charge and amine density of the tag, characteristics which were determined by the tag composition. The six lysine tag (Lys6) was found to be the most efficient (Allard, L., et al. (2001) Bioconjugate Chem. 12, 972). In the present work, the biological activity of Lys6-proteins covalently bound to polymer was investigated. N- or C-terminal Lys6-tagged HIV-1 p24 recombinant proteins (RK24H and RH24K) were grafted onto MAMVE, and the antigenicity each of the bioconjugates was evaluated using six monoclonal antibodies that recognized different epitopes distributed along the protein. We demonstrate that the position of the tag and the hydrolysis rate of the anhydride moieties of the polymer are the two main parameters involved in the conservation of the biological activity of the immobilized protein. We thus present a process which allows an efficient oriented immobilization of proteins onto copolymers with optimal biological activity that is suitable for the controlled production of active bioconjugates. |
| Databáze: | OpenAIRE |
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