کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2034029 | 1071985 | 2012 | 7 صفحه PDF | دانلود رایگان |

This study involved the fractionation of equine antivenom F(ab′)2 by combined stepwise ammonium sulfate (AS) and caprylic acid (CA) precipitation without intermediate separation of precipitate. Using a microplate format, 55 conditions with combinations of AS (0–20% saturation) and CA (0–5% v/v), were tested. AS significantly reduced the turbidity raised by CA. High specific antibody activity was observed in the area containing 2–5% CA and 10–20% AS. From these results, 12 precipitation conditions were selected for detailed quantitative studies. Two combinations, one with 4% CA and 15% AS and another with 5% CA and 20% AS, gave the highest fold-purification (1.79 and 1.83) with antibody recoveries at 68% and 59%, respectively. These combinations offered a benefit over CA alone in reducing the turbidity and in increasing the purity but not the recovery of antibody. The conditions giving more favorable overall results were with 2% CA alone and another with a combination of 1.5% CA and 10% AS. These preparations of F(ab′)2 were homogeneous and without protein aggregate under size-exclusion HPLC. Lastly, 1 h precipitation showed better results than those of overnight precipitation. These results could be useful for the production of therapeutic antivenoms.
► Antivenom F(ab′)2 was fractionated by combinations of ammonium sulfate and caprylic acid.
► The effects of precipitants on turbidity, purity and recovery of F(ab′)2 were investigated.
► Optimum fractionation conditions for F(ab′)2 were proposed.
► Better overall results were observed with 1 h than with overnight precipitation.
Journal: Biologicals - Volume 40, Issue 5, September 2012, Pages 338–344