A Platform for SpyCatcher Conjugation to Native Antibodies DOI Creative Commons
Soňa Krajčovičová, Thomas Wharton, C. Driscoll

et al.

Chemical Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Protein-antibody conjugates represent major advancements in targeted therapeutics. However, platforms enabling 'off-the-shelf' antibody conjugation are seldom reported. The SpyTag/SpyCatcher system, known for its stable isopeptide bond formation, is widely used to engineer protein architectures and study folding. This work introduces the fusion of SpyCatcher with native antibodies using cysteine-reactive tetra-divinylpyrimidine (TetraDVP)-SpyTag linkers. platform allows rapid a proteins. As proof concept, HER2-targeting trastuzumab was conjugated different proteins TetraDVP-SpyTag linker, producing robust that retained specific binding HER2-positive cells excellent conversion rates. To demonstrate platform's broader applicability, linker successfully additional IgG1 IgG4 (durvalumab, brentuximab, cetuximab, gemtuzumab) similarly high efficiency as trastuzumab. Moreover, scalable solid-phase synthesis TetraDVP linkers has been developed, achieving yields purity. innovative enables precise, single-step bioconjugation, offering strong potential protein-antibody conjugate synthesis. With applications across therapeutics diagnostics, this method advances antibody-based drug development.

Language: Английский

A Platform for SpyCatcher Conjugation to Native Antibodies DOI Creative Commons
Soňa Krajčovičová, Thomas Wharton, C. Driscoll

et al.

Chemical Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Protein-antibody conjugates represent major advancements in targeted therapeutics. However, platforms enabling 'off-the-shelf' antibody conjugation are seldom reported. The SpyTag/SpyCatcher system, known for its stable isopeptide bond formation, is widely used to engineer protein architectures and study folding. This work introduces the fusion of SpyCatcher with native antibodies using cysteine-reactive tetra-divinylpyrimidine (TetraDVP)-SpyTag linkers. platform allows rapid a proteins. As proof concept, HER2-targeting trastuzumab was conjugated different proteins TetraDVP-SpyTag linker, producing robust that retained specific binding HER2-positive cells excellent conversion rates. To demonstrate platform's broader applicability, linker successfully additional IgG1 IgG4 (durvalumab, brentuximab, cetuximab, gemtuzumab) similarly high efficiency as trastuzumab. Moreover, scalable solid-phase synthesis TetraDVP linkers has been developed, achieving yields purity. innovative enables precise, single-step bioconjugation, offering strong potential protein-antibody conjugate synthesis. With applications across therapeutics diagnostics, this method advances antibody-based drug development.

Language: Английский

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