Cytoplasmic versus periplasmic expression of site-specifically and bioorthogonally functionalized nanobodies using expressed protein ligation
نویسندگان
چکیده
منابع مشابه
An efficient protocol towards site-specifically clickable nanobodies in high yield: cytoplasmic expression in Escherichia coli combined with intein-mediated protein ligation.
In this study, several expression strategies were investigated in order to develop a generic, highly productive and efficient protocol to produce nanobodies modified with a clickable alkyne function at their C-terminus via the intein-mediated protein ligation (IPL) technique. Hereto, the nanobody targeting the vascular cell adhesion molecule 1 (NbVCAM1) was used as a workhorse. The highlights o...
متن کاملStreamlined Expressed Protein Ligation Using Split Inteins
Chemically modified proteins are invaluable tools for studying the molecular details of biological processes, and they also hold great potential as new therapeutic agents. Several methods have been developed for the site-specific modification of proteins, one of the most widely used being expressed protein ligation (EPL) in which a recombinant α-thioester is ligated to an N-terminal Cys-contain...
متن کاملIMPROVEMENT OF hGM-CSF EXPRESSION USING A FUSION SYSTEM BY DIRECTING PROTEIN TO THE PERIPLASMIC SPACE
Bacterial expression systems provide economic and logistic advantages in manufacturing proteins for human therapeutic purposes. However, most such proteins accumulate in insoluble biologicaIIy inactive form when overexpressed in bacterial cells. This is the case while attempting to produce recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) in E. coli. In this repor...
متن کاملExpressed protein ligation
The introduction of noncanonical amino acids and biophysical probes into peptides and proteins, and total or segmental isotopic labelling has the potential to greatly aid thedeterminationofprotein structure, functionandprotein– protein interactions. To obtain a peptide as large as possible by solid-phase peptide synthesis, native chemical ligation was introduced to enable synthesis of proteins ...
متن کاملhuman tissue plasminogen activator expression in escherichia coli using cytoplasmic and periplasmic cumulative power
tissue plasminogen activator (tpa) is a serine protease, which is composed of five distinct structural domains with 17 disulfide bonds, representing a model of high-disulfide proteins in human body. one of the most important limitations for high yield heterologous protein production in escherichia coli (e. coli) is the expression of complex proteins with multiple disulfide bridges. in this stud...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Protein Expression and Purification
سال: 2017
ISSN: 1046-5928
DOI: 10.1016/j.pep.2017.02.009