摘要
1969年,Hadding等首次发表了从人血浆中纯化C_9的方法,但该法步骤复杂,回收率只有0.3％左右。1980年,Biesecker等报道一种改进的方法,即采用聚乙二醇(PEG)沉淀,L-Lys-Sepha-rose亲和层析去除纤溶酶原(pg),DEAE-Sephadex柱层析和羟基磷灰石(HA)柱层析4个步骤,得到纯化的人C_9,使C_9回收率得以提高。本实验参考Biesecker等方法分离C_9,摸索和改进层析条件,对HA的制备方法进行改良,从而大大提高了HA柱的流速和分离效果,得到回收率高,高纯化而活性保持完整的人C_9,为探讨红细胞膜上自身限制因子的功能活性及末端补体复合物新抗原研究奠定了基础。
A large amount of human C9 was purified from plasma by the following procedures; 1) Polyethylene glycol precipitation; 2) Depletion of plasminogen by passing over an L-lys-sepharose column; 3) DEAE-sephadex A-50 chro-matography; and 4) Hydroxylapatite (HA) chromatography.The method of C9 purification was improved by altering the column-elution conditions and by the establishment of a novel method for preparing high-flow-rate HA. As a result, the rate of recovery of C9 was high (28.2%) and no impurities were detected either on gel electrophoretic or immunochemical examination. The hemolytic activity of purified C9 was retained.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1992年第5期389-393,共5页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金资助课题
