Study on separation and purification process of C1 esterase inhibitor
Journal Title: Chinese Journal of Blood Transfusion - Year 2022, Vol 35, Issue 11
Abstract
Objective To study the technology of separating and purifying C1 esterase inhibitor (C1-INH) by using the waste washing liquid as raw materia during the preparation of human prothrombin complex (PCC) l. Methods C1-INH was isolated and purified by a two-step method of polyethylene glycol (PEG) 4000 precipitation and cation chromatography. The pH of raw materials and the concentration of PEG4000 were adjusted to investigate the optimal conditions of PEG4000 precipitation method. After PEG was precipitated and centrifuged, the supernatant is treated as the loading solution for cation exchange chromatography, using Fractogel EMD SE HiCap(M) gel and CM Sepharose FF gel for ion exchange chromatography. The most suitable gel and separation conditions were selected by comparing the C1-INH antigen yield, activity yield and specific activity. Results Under the condition of pH 6.1, when the mass fraction of PEG4000 was 14%, the recovery rate of C1 esterase inhibitor was close to 70%, and the removal rate of ceruloplasmin was more than 95% after stirring for 10 minutes. As fractogel EMD SE HiCap(M) gel was used for cation exchange chromatography, when the eluent salt concentration was 0.25 M sodium chloride, the activity yield of C1 esterase inhibitor was greater than 80%, and the specific activity was greater than 5 IU/mg. Conclusions Using the waste washing liquid as the raw material during the preparation of PCC, the C1 esterase inhibitor with high specific activity can be prepared through PEG precipitation and purification by Fractogel EMD SE HiCap(M) ion exchange chromatography.
Authors and Affiliations
Jiabin XU, Yawen LI, Jiali GONG, Erhua LUO, Meng ZHANG
Keywords
Related Articles
- EP ID EP738402
- DOI 10.13303/j.cjbt.issn.1004-549x.2022.11.007
- Views 4
- Downloads 0
Research progress of platelets and their derivatives based drug delivery systems
Platelets play an important role in physiological and pathological activities such as thrombosis, inflammation and tumorigenesis. At present, the application of platelets in drug delivery systems is increasingly studied....
Genetic background of RhD negative blood donors
Objective To analyze the genetic background of RhD-negative blood donors by detecting RHD and RHCE genes of those donors. Methods From March 2021 to May 2022, the blood samples of RhD-negative blood donors, who had been...
Matching strategy for patients with platelet transfusion refractoriness caused by compound antibodies against HLA and CD36
Objective To search compatible and suitable platelets for platelet transfusion refractoriness (PTR) patient caused by compound antibodies against HLA and CD36. Methods ELISA method was used to detect the antibody against...
Clinical characteristics and transfusion strategies of delayed serological transfusion reactions caused by platelet transfusion in tumor patients
Objective To analyze the clinical manifestations of delayed serological transfusion reactions (DSTR) after platelet transfusion in tumor patients, and to explore the transfusion strategy. Methods Clinical data and labora...
Factors affecting the efficacy of platelet transfusion and specificity of HLA-Ⅰ antibodies in tumor patients
Objective To analyze the factors affecting the efficacy of platelet transfusion and the specificity of HLA-Ⅰ antibodies in tumor patients. Methods Tumer patients applied for platelet transfusion from November 2019 to Jul...