Study On Purification Technology Of Plasma Protein

Ion exchange chromatography （IEC） is a simple, rapid, effective technique usedin protein separation, that mainly depends on different ability of various proteins to beadsorbed onto the adsorbent, which has advantages of high capacity and low cost.Animal plasma （serum） contains abundant proteins, and it is important to maintain thebody’s normal physiological activities. This methods of immunoglobulin, albumin andantithrombin purification in plasma has been built through Q Sepharose^TM–XL stronganion exchange chromatography coupled with molecular exclusion chromatography.The isoelectric point of immunoglobulin （Ig） and serum albumin （SA） in animalserum are about7.8and4.8, respectively. Based on their larger difference ofisoelectric point, and the pH gradient elution was performed, Q Sepharose^TM–XLstrong anion exchange chromatography coupled with molecular exclusionchromatography were used to purify Ig and SA simultaneously from the high immunerabbit serum. The results of protein content showed that the purification recoveries ofIg and SA were over95%and over90%, respectively. The method has the advantagesof simple operation and rapidity, and Ig and SA which could be purifiedsimultaneously from the animal serum maintain normal activities.New Zealand white rabbit was immunized using arginine kinase purchased bysigma company, Q SephroseTM-XL strong anion exchange chromatography was usedto purify arginine kinase polyclonal antibody from serum. The purity and activity ofarginine kinase polyclonal antibody were determined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis（SDS-PAGE） and indirect enzyme linked immunemethod, respectively．In conclusion, purified antibody showed high purity and normalactivity, and which has far-reaching significance in the aspect of polyclonal antibody purification in the future.Plasma antithrombin is the most important anti-clotting substance in the body，generally maintained at150mg/L²00mg/L levels. This study of antithrombinpurification was to applied in the plasma, respectively by Q Sepharose^TM–XL stronganion exchange chromatography and Heparin sodium-Sepharose-4B affinitychromatography. The protein molecular weight and purity were identified bySDS-PAGE, The present results showed that the molecular weight was about58KDathat was consistent with the standard antithrombin sample, meanwhile the purity isrelatively high.