Abstract
We tried to prepare HBV (hepatitis B virus)-free modified hemoglobin (PEG-PLP-Hb) solutions from HBV positive blood. The amount of HBV-DNA and HB antigens were monitored at each washing process and preparation step of PEG-PLP-Hb. The HBV markers were strikingly decreased after washing red cells four times with physiological saline at 1, 700×g for 10min. Although hemolysate contained a small amount of HBV-DNA, more than 99 percent of HBV-DNA were washed out. A residual amount of HBV-DNA left in hemolysate became below the detectable level at the SFH (stroma-free hemoglobin) preparation. In order to obtain a safer SFH preparation, we tested whether Dane particles can be removed by a micro-porous cellulose hollow fiber, BMM (Bemberg Microporous Membrane). A SFH preparation containing Dane particles was filtrated through the BMM-30 (30nm pore size). More than 86 percent of proteins were recovered in the filtrated fractions, whereas Dane particles were left in the concentrated fraction and became below detectable level in the filtrated fractions. Dane particles can be effectively removed by the BMM filter to less than 0.1 percent of the initial amount. The results seem to indicate that contamination of Dane particles in hemoglobin preparations can be avoided to a considerable degree by repeated red cell washing, removal of stroma and BMM filtration of hemoglobin preparation.
