Publications by authors named "B Billen"
Article Synopsis
- An intein-mediated protein ligation (IPL) method is introduced for producing nanobodies with a clickable alkyne group at their C-terminus, enhancing their functionality.
- The process involves expressing and extracting a nanobody-intein-chitin binding domain fusion protein in E. coli SHuffle T7 cells using LB medium, resulting in high yields of alkynated nanobodies that maintain their target-binding ability.
- This efficient and cost-effective protocol enables the large-scale production of modified nanobodies for applications in biosurfaces and advanced medical techniques like targeted drug delivery and imaging surgery through covalent coupling with azide-containing agents.
Given the major structural role phosphodiesters play in the organism it is surprising they have not been more widely adopted as a building block in sophisticated biomimetic hydrogels and other biomaterials. The potential benefits are substantial: phosphoester-based materials show excellent compatibility with blood, cells, and a remarkable resistance to protein adsorption that may trigger a foreign-body response. In this work, a novel class of phosphodiester-based ionic hydrogels is presented which are crosslinked via a phosphodiester moiety.
View Article and Find Full Text PDFSite-specific functionalization of nanobodies after introducing bioorthogonal groups offers the possibility to biofunctionalize surfaces with a uniformly oriented layer of nanobodies. In this paper, expressed protein ligation (EPL) was used for site-specific alkynation of the model nanobody NbBcII10. In contrast to EPL constructs, which are typically expressed in the cytoplasm, nanobodies are expressed in the periplasm where its oxidizing environment ensures a correct folding and disulfide bond formation.
View Article and Find Full Text PDFIn 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 of the protocol can be ascribed to a cytoplasmic expression of the nanobody-intein-chitin-binding domain fusion protein in the Escherichia coli SHuffle(®) T7 cells with a C-terminal extension, i.
View Article and Find Full Text PDFArticle Synopsis
- TRPV3 is a thermosensitive ion channel found in the skin, nose, and tongue, playing a role in sensing temperature, pain in inflamed tissues, skin sensitivity, and hair growth.
- Current research on TRP channels enhances our understanding of the mechanisms of pain and temperature sensation, but their molecular workings remain complex and poorly understood.
- This study introduces a new method for purifying the human TRPV3 protein, demonstrating its functionality and revealing how different ligands interact with the channel, paving the way for more detailed structural and functional investigations.