Efficient Phage Display with Multiple Distinct Non-Canonical Amino Acids Using Orthogonal Ribosome-Mediated Genetic Code Expansion

Benjamí Oller-Salvia, Jason W. Chin

Producció científica: Article en revista indexadaArticleAvaluat per experts

39 Cites (Scopus)

Resum

Phage display is a powerful approach for evolving proteins and peptides with new functions, but the properties of the molecules that can be evolved are limited by the chemical diversity encoded. Herein, we report a system for incorporating non-canonical amino acids (ncAAs) into proteins displayed on phage using the pyrrolysyl-tRNA synthetase/tRNA pair. We improve the efficiency of ncAA incorporation using an evolved orthogonal ribosome (riboQ1), and encode a cyclopropene-containing ncAA (CypK) at diverse sites on a displayed single-chain antibody variable fragment (ScFv), in response to amber and quadruplet codons. CypK and an alkyne-containing ncAA are incorporated at distinct sites, enabling the double labeling of ScFv with distinct probes, through mutually orthogonal reactions, in a one-pot procedure. These advances expand the number of functionalities that can be encoded on phage-displayed proteins and provide a foundation to further expand the scope of phage display applications.

Idioma originalAnglès
Pàgines (de-a)10844-10848
Nombre de pàgines5
RevistaAngewandte Chemie - International Edition
Volum58
Número32
DOIs
Estat de la publicacióPublicada - 5 d’ag. 2019
Publicat externament

Fingerprint

Navegar pels temes de recerca de 'Efficient Phage Display with Multiple Distinct Non-Canonical Amino Acids Using Orthogonal Ribosome-Mediated Genetic Code Expansion'. Junts formen un fingerprint únic.

Com citar-ho