Stanker L H, Vanderlaan M, Juarez-Salinas H
J Immunol Methods. 1985 Jan 21;76(1):157-69. doi: 10.1016/0022-1759(85)90488-0.
A single-step method for purification of mouse monoclonal antibodies directly from ascitic fluids using hydroxylapatite column chromatography is described. The procedure yields highly purified IgG or IgM antibodies. The purified immunoglobulin is essentially free of contaminating mouse albumin, transferrin, and other ascites proteins, as determined by SDS-polyacrylamide gel electrophoresis. Hydroxylapatite chromatography can also separate monoclonal IgG antibodies from contaminating IgG antibodies found in ascites fluid of animals that have been immunosuppressed prior to ascites induction. Furthermore, the evidence presented here suggests that some hybridomas of SP2/0 origin synthesize an extraneous light chain resulting in the secretion of hybrid antibody molecules.
描述了一种使用羟基磷灰石柱色谱法直接从腹水液中纯化小鼠单克隆抗体的单步方法。该方法可产生高度纯化的IgG或IgM抗体。通过SDS-聚丙烯酰胺凝胶电泳测定，纯化的免疫球蛋白基本不含污染的小鼠白蛋白、转铁蛋白和其他腹水蛋白。羟基磷灰石色谱法还可以从腹水诱导前接受免疫抑制的动物腹水中发现的污染性IgG抗体中分离出单克隆IgG抗体。此外，此处提供的证据表明，一些SP2/0来源的杂交瘤合成了一种外来轻链，导致杂交抗体分子的分泌。
