Synthetic Tuning of Domain Stoichiometry in Nanobody-Enzyme Megamolecules.
Synthetic Tuning of Domain Stoichiometry in Nanobody-Enzyme Megamolecules.
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DOI：
10.1021/acs.bioconjchem.0c00578
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发表时间：
2021-01-20
影响因子：
4.7
通讯作者：
Mrksich M
中科院分区：
文献类型：
--
作者：
Metcalf KJ;Kimmel BR;Sykora DJ;Modica JA;Parker KA;Berens E;Dai R;Dravid VP;Werb Z;Mrksich M
This paper presents a method to synthetically tune atomically precise megamolecule nanobody–enzyme conjugates for prodrug cancer therapy. Previous efforts to create heterobifunctional protein conjugates suffered from heterogeneity in domain stoichiometry, which in part led to the failure of antibody–enzyme conjugates in clinical trials. We used the megamolecule approach to synthesize anti-HER2 nanobody–cytosine deaminase conjugates with tunable numbers of nanobody and enzyme domains in a single, covalent molecule. Linking two nanobody domains to one enzyme domain improved avidity to a human cancer cell line by 4-fold but did not increase cytotoxicity significantly due to lowered enzyme activity. In contrast, a megamolecule composed of one nanobody and two enzyme domains resulted in an 8-fold improvement in the catalytic efficiency and increased the cytotoxic effect by over 5-fold in spheroid culture, indicating that the multimeric structure allowed for an increase in local drug activation. Our work demonstrates that the megamolecule strategy can be used to study structure–function relationships of protein conjugate therapeutics with synthetic control of protein domain stoichiometry.
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影响因子：
4.7
作者：
Agarwal, Paresh;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
15
作者：
Kim CH;Axup JY;Dubrovska A;Kazane SA;Hutchins BA;Wold ED;Smider VV;Schultz PG
通讯作者：
Schultz PG
DOI：
10.1073/pnas.072685299
发表时间：
2002-04-16
影响因子：
11.1
作者：
Hodneland, CD;Lee, YS;Mrksich, M
通讯作者：
Mrksich, M
影响因子：
4.8
作者：
Kumar, Ramasamy P.;Morehouse, Benjamin R.;Oprian, Daniel D.
通讯作者：
Oprian, Daniel D.
影响因子：
3.7
作者：
Kute, T;Lack, CM;Vaughn, JP
通讯作者：
Vaughn, JP