Suomela H, Myllylä G, Raaska E
Vox Sang. 1977 Jul;33(1):37-50. doi: 10.1111/j.1423-0410.1977.tb02234.x.
A method for the large-scale preparation of a coagulation factor IX concentrate from human plasma is described. The method includes absorption of the coagulation factors from cryosupernatant plasma to DEAE-Sephadex, extensive washing of the gel and elution of the coagulation factors with 0.5 M phosphate buffer at 6.85, followed by desalting of the eluate in a column of Sephadex G-25, then by lyophilization, dissolving and sterile filtration, and finally by freeze-drying of the final product. Experiments performed with HBsAg-positive plasma demonstrated a decrease in the HBsAg content by a factor of 10(-5) during the process. The product is inactive in a Na-PTT assay. The process yields an about 100-fold purified factor IX concentrate containing also factors II, VII, and X, but in relatively smaller amounts. The average yield relative to factor IX is 60%. The batch size has been from 16 to 150 litres of plasma and about 300 batches of the concentrate have been prepared. About 5,100 bottle (about 4.0 X 10(6) U of factor IX) of the concentrate have been used in the treatment of patients with haemophilia B. The clinical effect has always been good and the in vivo response to factor IX was 1.15 +/- 0.30%/U/kg body weight. Two cases of HBsAg-negative hepatitis that may have been caused by the concentrate, were detected. No thrombotic complications were found.
描述了一种从人血浆大规模制备凝血因子IX浓缩物的方法。该方法包括将凝血因子从冷上清血浆吸附到DEAE-葡聚糖上，对凝胶进行充分洗涤，并用0.5M磷酸盐缓冲液（pH 6.85）洗脱凝血因子，然后在葡聚糖G-25柱上对洗脱液进行脱盐，接着进行冻干、溶解和无菌过滤，最后对最终产品进行冷冻干燥。用HBsAg阳性血浆进行的实验表明，在此过程中HBsAg含量降低了10^(-5)倍。该产品在钠-部分凝血活酶时间（Na-PTT）测定中无活性。该方法得到一种约100倍纯化的凝血因子IX浓缩物，其中还含有凝血因子II、VII和X，但含量相对较少。相对于凝血因子IX的平均产率为60%。批次规模为16至150升血浆，已制备了约300批浓缩物。约5100瓶（约4.0×10^6单位的凝血因子IX）浓缩物已用于治疗乙型血友病患者。临床效果一直良好，凝血因子IX的体内反应为1.15±0.30%/单位/千克体重。检测到两例可能由该浓缩物引起的HBsAg阴性肝炎病例。未发现血栓形成并发症。
