| Prothrombin Complex Concentration(PCC), which has always been used for a replacement therapy in hemophilia B patients, were consisted of FⅡ,Ⅶ,ⅨandⅩ, all of which were synthesized in liver and were called Vitamin K dependent(VKD). Now, the production of PCC was made through the gel resin such as DEAE Sephadex A50 or other types, which have low mechanical strength and much losing during the preparation process. In addition, the batch procedure, which used in the preparation of PCC, has the problems of small scale, troublesome operation, virus infection and the problem of biological dangerous to the operators, all of which lead to the high cost of PCC and prevent its clinical use. Therefore, it is significant to find a novel ion exchanger which will be cheaper than the gels and suitable for the seal hermetical and continual operation, which aim to lower the cost of preparation of PCC.In this paper, because of its better adsorption behavior, the CG-6 was screened from a series of macro-ion exchangers through the static experiments in which the adsorption quantity and velocity were evaluated. Then the structure and some properties were tested. Based on studies about the adsorption equilibrium and adsorption characterization for PCC, the adsorption thermodynamics and kinetics were evaluated; The effect of different factors about breakthrough curves of PCC on CG-6 fixed bed were also tested. At last, the isolation and purification of PCC from the plasma were researched. The result showed that: The CG-6, screened out from the static experiment in which the adsorption quantity reached 65.337mg/g and the adsorption rate was over 87%, has the average particle diameter of 165. 89μm and average pore diameter of 153.57 nm, which indicated the classic macroporous structure.The result from the adsorption characterization showed that the phosphoric buffer is suitable for the PCC adsorption on CG-6 and adsorption quantity could reach 117.65mg/g.The adsorption equilibrium of PCC on CG-6 could described by Langmuir model and the enthalpy was 115.1kJ/mol, the entropy is 2.37(15℃),20.97(20℃),23.11(25℃),37.32(30℃) kJ·mol-1·K-1, and the free energy is-0.54(15℃),-5.93(20℃),-6.66(25℃),-11.20(30℃)kJ·mol-1. The kinetics of adsorption was plotted by pseudo second order model and the adsorption velocity parameter was 0.00103g/mg·min.Through testing the adsorption breakthrough behavior of PCC on CG-6 in the fixed bed, the results showed that lowing the flow velocity and PCC concentration was helpful to increase the adsorption rate of PCC solution, which was increased from51.45% to 73.98% and 32.14% to 61.64%. But the utilization ratio of CG-6 resin would be decrease if the H0/φwas increased continually. The results of testing the pH of the breakthrough solution indicated that it was not the macroporous structure but the ion exchange process for result of the adsorption mechanism research.During the preliminary study on PCC isolation and purification from plasma, the activity of the elution were evaluated, and the result showed that much of the PCC was adsorbed by CG-6, which indicated good adsorption performance of CG-6 in plasma. PCC could be eluted by the 0.8mol/l and 1.2mol/l NaCl, 0.01mol/L PB solution, the specific activity was 0.383 and 1.08IU/mg, which was 40 and 108 time purification factors compared to the plasma.As a novel macroporous ion exchanger, CG-6 had good protein adsorption behavior and could be used as a type of protein exchanger. Through the preliminary study on PCC isolation and purification with CG-6, it could be known that CG-6 has good adsorption characterization for PCC in plasma in terms of high specific activity and purification time. So, the further research about use of CG-6 in the new preparation method for PCC could be done in the next work.
|
PDF Full Text Request