The anti-angiogenic, carboxy terminal non-collagenous domain (NC1) derived from human Collagen type IV
alpha 6 chain, [α6(IV)NC1] or hexastatin, was earlier obtained using different recombinant methods of expression in bacterial
systems. However, the effect of L-arginine mediated renaturation in enhancing the relative yields of this protein
from bacterial inclusion bodies has not been evaluated. In the present study, direct stirring and on-column renaturation
methods using L-arginine and different size exclusion chromatography matrices were applied for enhancing the solubility
in purifying the recombinant α6(IV)NC1 from bacterial inclusion bodies. This methodology enabled purification of higher
quantities of soluble protein from inclusion bodies, which inhibited endothelial cell proliferation, migration and tube formation.
Thus, the scope for L-arginine mediated renaturation in obtaining higher yields of soluble, biologically active
NC1 domain from bacterial inclusion bodies was evaluated.
Keywords: Non-collagenous domain of α6 type IV collagen, L-arginine mediated renaturation, size exclusion chromatography, anti-angiogenic activity, human umbilical vein endothelial cells and vascular endothelial growth factor, inclusion bodies, blood capillaries
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