小鼠腹水IgM型单抗的纯化

Purification of IgM monoclonal antibodies from mouse ascitesfluid

目的：为了建立一种从腹水中获得高纯度ＩｇＭ 抗体的纯化方法。方法：分别以硫酸铵沉淀或优球蛋白沉淀结合凝胶过滤法，从腹水中纯化ＩｇＭ 型单克隆抗体。结果：用优球蛋白沉淀结合凝胶过滤，纯化的ＩｇＭ 抗体纯度＞ ９０％ ，明显优于硫酸铵沉淀结合凝胶过滤法纯化的ＩｇＭ 抗体纯度（ ＜６０％ ）。结论：优球蛋白沉淀结合凝胶过滤是一种获得高纯度ＩｇＭ 抗体的方法。

Objective:To obtain high purity and immunoreactive IgMantibodyfrom ascites. Methods:Ascitesfluid was subjected to ammonium sulfate precipitation oreuglobulin precipitation.The precipitate wasredissolved and subsequently subjectedto gelfiltration on Su perdexTM 200 .The purity ofthe antibody wasassessed bythereducing SDSgelelectrophoresis.Immuno activity ofthe antibody wasassessed byanindirectenzyme linkedimmunosorbentassay.Results:The purity ofthe antibody wasestimatedto begreaterthan 90% aftereuglobulin precipitation combined with gelfiltration,butlessthan 60 % afterammonium sulfate precipitationcombined withgelfiltration.Thereisnoob viously difference between two method’s recovery ofimmuno activity. Conclusion :Euglobulin precipitation combined with gelfiltration is a good methodfor purificationIgM Mabsfrom ascites.