Open AccessCell-free synthesis of human toll-like receptor 9 (TLR9): Optimization of synthesis conditions and functional analysis
Author(s) -
Srujan Kumar Dondapati,
Georg Pietruschka,
Lena Thoring,
Doreen A. Wüstenhagen,
Stefan Kubick
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0215897
Subject(s) - ectodomain , tlr9 , receptor , innate immune system , toll like receptor 9 , pathogen associated molecular pattern , toll like receptor , biology , microbiology and biotechnology , immunology , computational biology , pattern recognition receptor , gene , genetics , gene expression , dna methylation
The Toll-like receptor family belongs to the group of pathogen recognition receptors which is responsible for the discrimination of self and non-self pathogen-associated molecular patterns (PAMP’s). Toll-like receptors play an important role in the innate immunity and defects in protein expression or polymorphism is linked to various diseases such as Systemic Lupus Erythematosus (SLE). The elucidation of the underlying mechanism is crucial for future treatment and therapeutics of toll-like receptor linked diseases. Herein, we report the cell-free synthesis of human Toll-like receptor 9 (hTLR9) using CHO lysate and the continuous exchange cell-free (CECF) synthesis platform. The functionality of this protein was demonstrated by an ELISA binding assay using the ectodomain of TLR9 (TLR9-ECD).