Tyrosinase-Mediated Synthesis of Nanobody-Cell Conjugates

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 nudeophilic 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.

Automated summary

A convenient enzymatic strategy is described for modifying cell surfaces using tyrosinase enzyme. The strategy involves introducing unique tyrosine residues at the C-termini of nanobodies, which can then be selectively oxidized to reactive o-quinones. These reactive intermediates readily react with thiol, amine, and imidazole residues present on cell surfaces, resulting in novel nanobody-cell conjugates that exhibit targeted antigen binding. The approach is further extended to the synthesis of nanobody-NK cell conjugates for targeted immunotherapy applications, which demonstrate targeted cell binding and induce targeted cell death.