VPB0228 - Investigation of properties of macropore silica sorbents and active dyes as ligands suitable for purification of factor VIII coagulation

Background: A number of synthetic dyes, as they react with a wide variety of biological materials could be used as affinity ligands. The commercial active dyes contain various impurities which may affect their biochemical and related use. The purification of FVIII is generally required for the treatment Haemophilia A or von Willebrand’s disease and heavy loss of blood, requires relatively high purity for medical use.

Aims: to investigate of properties macropore silica and active dyes as ligands.

Methods: chemical synthesis of macropore silica sorbents with active dyes by the method of "salt inclusion"; ultradiafiltration; spectrophotometric.

Results: We used in the work: Diasorb-aminopropyl 800/70, fraction 0.25–0.50 µm, Cibacron Brilliant Yellow 3GP, Procion Blue HB, Procion Blue MXR, Procion Yellow HE3G, Procion Gelb M4R, Procion Red MX5B, Reactive Brown 10, Reactive Red 120, Reactive Green 5, Reactive Green 19, Active bright orange KH, Active violet 4K, Active bright blue КН, Active bright red 5SН, Active burgundy 4SG and Active purple 4GT.
The chemical stability of sorbents was evaluated under the influence of high ionic strength of the solution, polar solvents, changes in pH and high temperature. The bond strength was assessed by detecting the free dye in the light-absorbing solution at the appropriate wavelength for each dye, which was measured on a DS-11FX spectrophotometer (DeNovix, England). The appearance of free dyes in the supernatant at high concentrations of NaCl (5.0 M) or isopropanol (50%) indicates that a small amount of dyes was bound to the matrix due to non-covalent interaction. The release of the dye into solution at pH 10.0 is due to the partial hydrolysis of the silica matrix.

Conclusion(s): Characteristics and advantages in practical application of affinity sorbents are determined.