Reference - Detail
| RRC ID | 87368 |
|---|---|
| Author | Agustriana E, Murozono K, Nishioka R, Kawaguchi Y, Kimura M, Kamiya N. |
| Title | Chemoenzymatic method for site-selective fluorophore conjugation of a native IgG Fab fragment. |
| Journal | J Biosci Bioeng |
| Abstract |
Antibody-derived bioconjugates have emerged as practical biomolecules for applications in diagnostics and therapeutics. A key challenge is achieving site-selective bioconjugation to preserve the native function of the labeled biomolecule. Methods that target native amino acid residues can broaden the applicability of bioconjugates; however, most existing methods rely on genetically engineered antibodies to ensure site-specific labeling. In this report, a chemoenzymatic strategy for modifying a native Fab fragment derived from trastuzumab was investigated using the EzMTG-pG fusion protein, which consists of a microbial transglutaminase variant and protein G. To mitigate the hydrophobic nature of the widely used dibenzocyclooctyne (DBCO) moiety for click chemistry, a new DBCO-containing glutamine donor peptide (DBCO-PEG4-LLQG) was designed. This substrate peptide enabled Lys65-selective conjugation of the Fab via EzMTG-pG catalysis, achieving a 92% modification rate within 4 h. The subsequent click reaction between the Fab-DBCO conjugate and an azide-bearing fluorescent small molecule probe generated a Fab-fluorophore conjugate that retained cell-specific binding to a target cell line. These results demonstrate that the chemoenzymatic pathway, using EzMTG-pG catalysis combined with a click reaction, provides a versatile approach for generating Fab-based bioconjugates with potential applications in diagnostics and therapeutics. |
| Published | 2026-4-8 |
| DOI | 10.1016/j.jbiosc.2026.03.005 |
| PII | S1389-1723(26)00121-0 |
| PMID | 41956928 |
| IF | 2.366 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 1.0 |
| Resource | |
| Human and Animal Cells | SK-BR-3(RCB2132) MDA-MB-231/luc(RCB5936) |