A Cell-Free Protein Expression System Derived from Human Primary Peripheral Blood Mononuclear Cells.
ACS Synth Biol
; 9(8): 2188-2196, 2020 08 21.
Article
em En
| MEDLINE
| ID: mdl-32698572
ABSTRACT
Historically, some of the first cell-free protein expression systems studied in vitro translation in various human blood cells. However, because of limited knowledge of eukaryotic translation and the advancement of cell line development, interest in these systems decreased. Eukaryotic translation is a complex system of factors that contribute to the overall translation of mRNA to produce proteins. The intracellular translateome of a cell can be modified by various factors and disease states, but it is impossible to individually measure all factors involved when there is no comprehensive understanding of eukaryotic translation. The present work outlines the use of a coupled transcription and translation cell-free protein expression system to produce recombinant proteins derived from human donor peripheral blood mononuclear cells (PBMCs) activated with phytohemagglutinin-M (PHA-M). The methods outlined here could result in tools to aid immunology, gene therapy, cell therapy, and synthetic biology research and provide a convenient and holistic method to study and assess the intracellular translation environment of primary immune cells.
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Base de dados:
MEDLINE
Assunto principal:
Leucócitos Mononucleares
/
Sistema Livre de Células
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article