Lipoate-acid ligase a modification of native antibody: Synthesis and conjugation site analysis
| Autor: | Inoue Kota, Yutaka Matsuda, Yasuhiro Mihara, Shikida Natsuki, Kazutoshi Takahashi, Shunsuke Yamazaki, Kazutaka Shimbo |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Immunoconjugates
Clinical Biochemistry Chemical biology Pharmaceutical Science Immunoglobulin light chain Biochemistry Ligases Drug Discovery Humans Molecular Biology chemistry.chemical_classification DNA ligase Bioconjugation Molecular Structure Thioctic Acid biology Organic Chemistry Fragment crystallizable region Enzyme chemistry Immunoglobulin G biology.protein Molecular Medicine Antibody Conjugate |
| Zdroj: | Bioorganic & Medicinal Chemistry Letters. 51:128360 |
| ISSN: | 0960-894X |
| DOI: | 10.1016/j.bmcl.2021.128360 |
| Popis: | Bioconjugation is an important chemical biology research focus, especially in the development of methods to produce pharmaceutical bioconjugates and antibody-drug conjugates (ADCs). In this report, an enzyme-catalyzed conjugation method combined with a chemical reaction was used to modify a native antibody under mild reaction conditions. Our investigation revealed that lipoic-acid ligase (LplA) modifies native IgG1 with biased site-specificity. An intact mass analysis revealed that 98.3% of IgG1 was modified by LplA and possessed at least one molecule of octanocic acid. The average number of modifications per antibody was calculated to be 4.6. Peptide mapping analysis revealed that the modified residues were K225, K249 and K363 in the Fc region, and K30, K76 and K136 in the heavy chain and K39/K42, K169, K188 and K190 in the light chain of the Fab region. Careful evaluation including solvent exposed amino acid analysis suggested that these conjugate sites were not only solvent exposed but also biased by the site-specificity of LplA. Furthermore, antibody fragment conjugation may be able to take advantage of this enzymatic approach. This feasibility study serves as a demonstration for preparing enzymatically modified antibodies with conjugation site analysis. |
| Databáze: | OpenAIRE |
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