Matches in Ghent University Academic Bibliography for { <https://biblio.ugent.be/publication/01GQ7VGA3N83P13VCV98P1R2PB> ?p ?o. }
Showing items 1 to 22 of
22
with 100 items per page.
- 01GQ7VGA3N83P13VCV98P1R2PB classification C3.
- 01GQ7VGA3N83P13VCV98P1R2PB date "2022".
- 01GQ7VGA3N83P13VCV98P1R2PB language "eng".
- 01GQ7VGA3N83P13VCV98P1R2PB type conference.
- 01GQ7VGA3N83P13VCV98P1R2PB hasPart 01GQ7VSW7HKXTCGFYSYYPHM3YK.pdf.
- 01GQ7VGA3N83P13VCV98P1R2PB subject "Technology and Engineering".
- 01GQ7VGA3N83P13VCV98P1R2PB subject "Veterinary Sciences".
- 01GQ7VGA3N83P13VCV98P1R2PB presentedAt urn:uuid:da1ea1a9-c67b-4da3-9ae0-3ba69b31959b.
- 01GQ7VGA3N83P13VCV98P1R2PB abstract "Introduction: Endolysins are a class of antimicrobials that are on the verge of break-through in (veterinary) medicine. Although the activity and engineering of these proteins is well-studied, insights regarding their stability, pharmaco-kinetics & -dynamics are limited in an eukaryotic cell environment or upon administration to either animals or humans. Objective: Our groups developed a method that allows the purification of non-pyrogenic endolysins purified from E. coli, applied to our previous research in the context of bovine mastitis. Materials, Methods and Results: ClearColi® BL21 (DE3), an E. coli line knocked-out for immunogenic lipopolysaccharide (LPS), was used for the expression of a polyhistidine (his)-tagged endolysin. After sonication on ice, the centrifuged lysate was applied to a HisTrap column on an Äkta Pure system, which was pretreated with 1M NaOH to disintegrate residual LPS. The column was washed with lysis buffer containing 0.1% EMPIGEN® detergent to remove any residual LPS left in the sample. Next, this detergent was removed from the column by washing with lysis buffer, after which his-rich host cell proteins were likewise removed with 50 mM imidazole. An isocratic elution with 400 mM imidazole yielded the his-tagged protein. Finally, buffer exchange was executed by centrifugation in a Pierce™ protein concentrator, also correcting the volume to the desired molarity. As a proof-of-concept for non-pyrogenicity, bovine mammary epithelial cells (boMECs) and a limited number of mouse mammary glands (n = 6) were incubated or intramammarily injected with 8 or 2.5 µM of purified endolysin, respectively. Neutral red staining evaluated cytotoxicity on the boMECs and histology on the mouse mammary glands also confirmed safety. Conclusion: Our method successfully purified one selected endolysin without precipitation or disintegration loss. This purified endolysin was regarded safe both in vitro and in vivo upon its administration to boMECs and mice.".
- 01GQ7VGA3N83P13VCV98P1R2PB author C157F57C-AA66-11E4-986E-18D2B4D1D7B1.
- 01GQ7VGA3N83P13VCV98P1R2PB author F4730512-F0ED-11E1-A9DE-61C894A0A6B4.
- 01GQ7VGA3N83P13VCV98P1R2PB author F4EEB47C-F0EE-11E1-A197-91C894A0A6B4.
- 01GQ7VGA3N83P13VCV98P1R2PB author F825BEB6-F0ED-11E1-A9DE-61C894A0A6B4.
- 01GQ7VGA3N83P13VCV98P1R2PB author f9b7dab4-027b-11ea-934d-87264d075fbf.
- 01GQ7VGA3N83P13VCV98P1R2PB author urn:uuid:d47517da-b485-4025-b7e5-c14e6659cbf1.
- 01GQ7VGA3N83P13VCV98P1R2PB dateCreated "2023-01-20T15:07:10Z".
- 01GQ7VGA3N83P13VCV98P1R2PB dateModified "2024-10-29T18:33:42Z".
- 01GQ7VGA3N83P13VCV98P1R2PB name "A method for the purification of non-pyrogenic bacteriophage derived endolysins with an in vitro or in vivo application".
- 01GQ7VGA3N83P13VCV98P1R2PB sameAs LU-01GQ7VGA3N83P13VCV98P1R2PB.
- 01GQ7VGA3N83P13VCV98P1R2PB sourceOrganization urn:uuid:73dd468c-c3cf-48f8-808d-f212c9d63609.
- 01GQ7VGA3N83P13VCV98P1R2PB sourceOrganization urn:uuid:c0fc38c8-1009-4404-bc0b-e52cab9b1b5b.
- 01GQ7VGA3N83P13VCV98P1R2PB type C3.