Purification of HBsAg from human placental blood by density-gradient zonal centrifugation

✍ Scribed by Yan-Bo Xie; Ji-Zhang Wang; Guo-Quiang Zhang; Li-Xin Wang

• Publisher: John Wiley and Sons
• Year: 1981
• Tongue: English
• Weight: 926 KB
• Volume: 7
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/jmv.1890070212

✦ Synopsis

Highly purified preparations of HBsAg are urgently needed for the production of hepatitis B diagnostic products and vaccines. The raw material for the production of HBsAg usually involves blood from HBsAg-positive carriers. In some countries, it is difficult to draw blood from such carriers due to legal or habitual reasons. Here we report the purification of HBsAg from HBsAg-positive human placental blood, which is considered a waste material in the routine screening of placenta for the production of placental blood derivatives.

MATERIALS A N D METHODS

Instruments

Hitachi 8OP-7 preparative ultracentrifuge and its RPZ zonal rotors. LKB Uvicord I1 ultraviolet monitor and fraction collector. Millipore ultrafiltration set.

Raw Material for Production of HBsAg

We used HBsAg-positive human placental blood as raw material. We collected placental blood serum with a CEP titer equal or above 1 :16 and kept it in a deep freezer below ~ 20°C. After thawing, we centrifuged the serum at 3,000 rpm for 20 minutes to remove the sediments prior to use.

Purification Procedures

As shown in the Results section we used two combinations of procedures for com-Ammonium sulfate precipitation. The ammonium sulfate precipitation was done Peptic digestion. This was performed according to the procedure given by Kim et parison. Here we give the details of each single procedure. at half saturation. The supernatant was discarded. a1 [ l ] .