Tyrosinase-Mediated Synthesis of Nanobody-Cell Conjugates

Johnathan C Maza; Derek M García-Almedina; Lydia E Boike; Noah X Hamlish; Daniel K Nomura; Matthew B Francis

doi:10.1021/acscentsci.1c01265

Tyrosinase-Mediated Synthesis of Nanobody-Cell Conjugates

ACS Cent Sci. 2022 Jul 27;8(7):955-962. doi: 10.1021/acscentsci.1c01265. Epub 2022 Jun 22.

Authors

Johnathan C Maza¹, Derek M García-Almedina¹, Lydia E Boike^{1

2}, Noah X Hamlish³, Daniel K Nomura^{1

2

3

4

5}, Matthew B Francis^{1

6}

Affiliations

¹ Department of Chemistry, University of California, Berkeley, California 94720, United States.
² Novartis-Berkeley Center for Proteomics and Chemistry Technologies, Cambridge, Massachusetts 02139, United States.
³ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California 94720, United States.
⁴ Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, California 94720, United States.
⁵ Innovative Genomics Institute, Berkeley, California 94720, United States.
⁶ Materials Sciences Division, Lawrence Berkeley National Laboratories, Berkeley, California 94720,United States.

Abstract

A convenient enzymatic strategy is reported for the modification of cell surfaces. Using a tyrosinase enzyme isolated from Agaricus bisporus, unique tyrosine residues introduced at the C-termini of nanobodies can be site-selectively oxidized to reactive o-quinones. These reactive intermediates undergo rapid modification with nucleophilic thiol, amine, and imidazole residues present on cell surfaces, producing novel nanobody-cell conjugates that display targeted antigen binding. We extend this approach toward the synthesis of nanobody-NK cell conjugates for targeted immunotherapy applications. The resulting NK cell conjugates exhibit targeted cell binding and elicit targeted cell death.