Solid-Phase Bioconjugation of Heterobifunctional Adaptors for Versatile Assembly of Bispecific Targeting Ligands
| Autor: | Pavel Zrazhevskiy, Hong Yan Liu, Xiaohu Gao |
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| Rok vydání: | 2014 |
| Předmět: |
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Molecular Streptavidin Protein Conformation Biomedical Engineering Pharmaceutical Science Bioengineering Nanotechnology Ligands 010402 general chemistry 01 natural sciences Article Polyethylene Glycols 03 medical and health sciences chemistry.chemical_compound Cell Line Tumor Antibodies Bispecific Humans Biotinylation Staphylococcal Protein A 030304 developmental biology Targeting ligands Pharmacology 0303 health sciences Bioconjugation Extramural Organic Chemistry Combinatorial chemistry 0104 chemical sciences Drug development chemistry Biotechnology |
| Zdroj: | Bioconjugate Chemistry |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc5002455 |
| Popis: | High-throughput generation of bispecific molecules promises to expedite the discovery of new molecular therapeutics and guide engineering of novel multifunctional constructs. However, high synthesis complexity and cost have hampered the discovery of bispecific molecules in drug development and biomedical research. Herein we describe a simple solid-phase bioconjugation procedure for preparation of Protein A(G,L)-PEG-Streptavidin heterobifunctional adaptors (with 1:1:1 stoichiometry), which enable self-assembly of unmodified antibodies and biotinylated molecules into bispecific targeting ligands in a versatile mix-and-use manner. Utility of such adaptors is demonstrated by assembly of anti-CD3 and anti-Her2 antibodies into bispecific CD3xHer2 targeting ligands, which efficiently drive T-cell-mediated lysis of Her2-positive cancer cells. In comparison to bioconjugation in solution, the solid-phase procedure described here offers precise stoichiometry control, ease of purification, and high yield of functional conjugates. Simplicity and versatility should prove this methodology instrumental for preparation of bispecific ligands, as well as for high-throughput screening of bispecific combinations, before proceeding to synthesis of lead candidates via recombinant engineering or chemical cross-linking. |
| Databáze: | OpenAIRE |
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