ABSTRACT
【Objective】 To optimize the purification conditions of heparin affinity chromatography in the purification of human coagulation factor Ⅸ by response surface method and establish the optimal chromatography process parameters. 【Methods】 The effect of sample loading temperature on purification efficiency was analyzed through single factor test. Three-factor three-level response surface method was used to optimize the chromatographic elution conditions. The Folin phenol method and the automatic hemagglutination analyzer were used to determine the total protein content and human coagulation factor Ⅸ titer, respectively. The purification effect was evaluated by activity index and process recovery rate. 【Results】 The optimized optimal chromatographic conditions were loading at 5 ℃, washing 4 CV, eluent formulation of sodium citrate 20 mmol/L, arginine hydrochloride 18.7 mmol/L, NaCl 611.6 mmol/L and pH 7.5; under this optimal setting, the recovery rate of the chromatographic process was (46.6±2.9) %, titer of factor Ⅸ rated to (68.4±4.7) IU/mL and specific activity was (62.8±3.3) IU/mg. 【Conclusion】 The optimized parameters of heparin affinity chromatography process by response surface method can produce better purification effect on human coagulation factor Ⅸ.
ABSTRACT
Objective To discusses the methods of the case selection and monitoring and diagnosis and treatment to selective non-operative management of closed splenic injuries .Methods The clinical data and results of 98 cases from October 1999 to Octo-ber 2011 with closed splenic injuries managed non-operative treatment were analyzed retrospectively .Results There were 90 cases (91 .8% ) conservative successfully ,5 cases(5 .1% ) transit operation because hematoma increase and bleeding continuously during therapeutic observation and 2 cases(3 .1% ) died from multiple injuries merge infection and multiple organ failure .Conclusion Most of the closed splenic injuries below level Ⅱcan be treated by non-operative management ,the effect and prognosis determined by the reasonable case selection and monitoring conditions and diagnosis and treatment .