Synthesis of Fluorescently Labeled Antibodies Using Non-Canonical Amino Acids in Eukaryotic Cell-Free Systems

Marlitt Stech, Nathanaël Rakotoarinoro, Tamara Teichmann, Anne Zemella, Lena Thoring, Stefan Kubick*

*Autor corresponent d’aquest treball

Producció científica: Capítol de llibreCapítolAvaluat per experts

6 Cites (Scopus)

Resum

Cell-free protein synthesis (CFPS) enables the development of antibody conjugates, such as fluorophore conjugates and antibody-drug conjugates (ADCs), in a rapid and straightforward manner. In the first part, we describe the cell-free synthesis of antibodies containing fluorescent non-canonical amino acids (ncaa) by using pre-charged tRNA. In the second part, we describe the cell-free synthesis of antibodies containing ncaa by using an orthogonal system, followed by the site-specific conjugation of the fluorescent dye DyLight 650-phosphine. The expression of the antibodies containing ncaa was analyzed by SDS-PAGE, followed by autoradiography and the labeling by in-gel fluorescence. Two different fluorescently labeled antibodies could be generated.

Idioma originalAnglès
Títol de la publicacióStructural Proteomics
Subtítol de la publicacióHigh-Throughput Methods
EditorsRaymond J. Owens
EditorHumana Press Inc.
Pàgines175-190
Nombre de pàgines16
ISBN (electrònic)978-1-0716-1406-8
ISBN (imprès)978-1-0716-1405-1
DOIs
Estat de la publicacióPublicada - 2021
Publicat externament

Sèrie de publicacions

NomMethods in Molecular Biology
Volum2305
ISSN (imprès)1064-3745
ISSN (electrònic)1940-6029

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