Purified human FVIII was electrophoresed through SDS agarose gels of different concentrations and through SDS Polyacrylamide (PA) gels of exceptionally large pore size (3.75% monomer, 1% cross linking). The gels were calibrated (a) with the FVIII subunit (b) with dimethyl-suberimidate cross-linked human fibrinogen. After Coomassie-blue staining the PA gels were scanned densito-metrically.The results show that freshly purified FVIII at pH 6.5 exists as a homologous series of oligomers and that a linear relationship obtains between the mobility of oligomers 1-6 and log molecular weight. Another linear relationship exists between the higher oligomers due to their significantly raised mobility. This suggests that the higher oligomers have a compact structure which is still partially ordered after SDS denaturation. A plot of log oligomer molarity vs. log subunit number has a gradient of approximately -1, suddenly decreasing to -5.5 after the hexamer, strongly indicating a decrease in the rate of polymerisation after this point. Oligomers 1-6 comprised 88% of the total FVIII, and oligomers 7-9 a further 6%.FVIII left at room temperature for 24 hours associated into extremely large oligomers.
Hydrogel Nanoparticles with Covalently Linked Coomassie Blue for Brain Tumor Delineation Visible to the Surgeon