摘要
采用一种经修改的辛酸沉淀法进行纯化小鼠腹水或血清中单克隆抗体(McAbs)的试验,全部八种抗促卵泡激素(FSH)单抗腹水的纯化结果都很好。该方法大致做法为将经稀释的腹水调pH至4.5后加入辛酸,离心弃沉淀,取上清液加等体积饱和硫酸铵,所得沉淀物用磷酸盐缓冲液(PBS)溶解后对相同PBS作广泛透析。以聚丙稀酰胺凝胶电泳(PAGE)检查经纯化抗体的纯度,并在600nm波长上扫描PAGE图谱,对抗体纯度作定量分析。结果抗体纯化物的平均纯度达93％(范围:83.5—98.6％)。通过酶联免疫吸附测定法(ELISA)检测得知,纯化后抗体活性无明显丢失。
Eight monoclonal antibodies (McAbs) to follicle-stimulating hormone (FSH) in murine ascitic fluids or sera are highly purified respectively using a modified caprylic acid precipitation method. Briefly, the pH of diluted ascitic fluid is adjusted to 4.5 and caprylic acid is added. The precipitate formed is discarded after centrifugation and supernatant is precipitated with an equal volume of saturated ammonium sulphate. The precipitate is then dissolved in PBS and dialyzed extensively against the same PBS buffer. The purity of each purified McAb is checked by polyacrylamide gel electrophoresis(PAGE) and the quantity is determined by gel-scanning at 600nm. The McAb solutions gave an average purity of 93% (range:83.5—98.6%). There are no significant losses in biological activities of all the eight purified McAbs as determined by enzyme-linked immunosorbent assay (ELISA).
出处
《河北省科学院学报》
CAS
1991年第2期56-61,共6页
Journal of The Hebei Academy of Sciences
关键词
单克隆抗体
辛酸沉淀
纯化
ELISA
(Monoclonal antibody) (Caprylic acid precipitation) (Purification of antibody)
