Direct capture and selective elution of a secreted polyglutamate-tagged nanobody using bare magnetic nanoparticles was written by Zanker, Alexander A.;Stargardt, Patrick;Kurzbach, Sophie C.;Turrina, Chiara;Mairhofer, Juergen;Schwaminger, Sebastian P.;Berensmeier, Sonja. And the article was included in Biotechnology Journal in 2022.Related Products of 367-93-1 The following contents are mentioned in the article:
The secretion and direct capture of proteins from the extracellular medium is a promising approach for purification, thus enabling integrated bioprocesses. We demonstrate the secretion of a nanobody (VHH) to the extracellular medium (EM) and its direct capture by bare, non-functionalized magnetic nanoparticles (MNPs). An ompA signal peptide for periplasmic localization, a polyglutamate-tag (E8) for selective MNP binding, and a factor Xa protease cleavage site were fused N-terminally to the nanobody. The extracellular production of the E8-VHH (36 mg L-1) was enabled using a growth-decoupled Escherichia coli-based expression system. The direct binding of E8-VHH to the bare magnetic nanoparticles was possible and could be drastically improved up to a yield of 88% by adding polyethylene glycol (PEG). The selectivity of the polyglutamate-tag enabled a selective elution of the E8-VHH from the bare MNPs while raising the concentration factor (5x) and purification factor (4x) significantly. Our studies clearly show that the unique combination of a growth-decoupled E. coli secretion system, the polyglutamate affinity tag, non-functionalized magnetic nanoparticles, and affinity magnetic precipitation is an innovative and novel way to capture and concentrate nanobodies. This study involved multiple reactions and reactants, such as (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1Related Products of 367-93-1).
(2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1) belongs to alcohols. Under appropriate conditions, inorganic acids also react with alcohols to form esters. To form these esters, a wide variety of specialized reagents and conditions can be used. The most common reactions of alcohols can be classified as oxidation, dehydration, substitution, esterification, and reactions of alkoxides.Related Products of 367-93-1
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