Affinity recovery of lentivirus by diaminopelargonic acid mediated desthiobiotin (cas 15720-25-9) labelling

Add time:09/03/2019    Source:sciencedirect.com

desthiobiotin (cas 15720-25-9)-tagged lentiviral vectors have been metabolically produced by DBL producer cells in a 7,8-diaminopelargonic acid (7-DAPA) dependent manner for envelope independent, single-step affinity purification. 7-DAPA, which has little or no affinity for avidin/streptavidin, was synthesised and verified by NMR spectroscopy and mass spectrometry. By expressing the biotin acceptor, biotin ligase and desthiobiotin synthase bioD, DBL cells converted exogenous 7-DAPA into membrane-bound desthiobiotin. Desthiobiotin on the DBL cell surface was visualised by confocal microscopy and the desthiobiotin density was quantified by HABA-avidin assay. Desthiobiotin was then spontaneously incorporated onto the surface of lentiviral vectors produced by the DBL cells. It has been demonstrated by flow cytometry that the desthiobiotinylated lentiviruses were captured from the crude 7-DAPA-containing viral supernatant by Streptavidin Magnespheres® and eluted by biotin solution efficiently whilst retaining infectivity. The practical, high yielding virus purification using Pierce monomeric avidin coated columns indicates a highly efficient biotin-dependent recovery of infectious lentiviruses at 68%. The recovered lentiviral vectors had a high purity and the majority were eluted within 45 min. This 7-DAPA mediated desthiobiotinylation technology can be applied in scalable production of viral vectors for clinical gene therapy.

We also recommend Trading Suppliers and Manufacturers of desthiobiotin (cas 15720-25-9). Pls Click Website Link as below: cas 15720-25-9 suppliers

Prev:Straightforward preparation of enantiopure 3-amino-4,4-dimethylpyrrolidin-2-one and its derivatives
Next:Easily reversible desthiobiotin (cas 15720-25-9) binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation)