Carty P, O'Kennedy R
School of Biological Sciences, National Institute for Higher Education, Dublin, Ireland.
J Chromatogr. 1988 Jun 17;442:279-88. doi: 10.1016/s0021-9673(00)94475-3.
The purification and characterization of polyclonal and monoclonal antibodies and antibody-enzyme conjugates by high-performance liquid chromatography (HPLC) using a Protein Pak 300 SW column and a Protein Pak DEAE anion-exchange column is described. The following polyclonal antibodies were examined: (i) donkey anti-mouse immunoglobulin G (IgG), (ii) mouse IgG, (iii) sheep anti-human IgG and (IV) goat anti-human factor VIII. HPLC was used to analyse the purity of horseradish peroxidase conjugates of rabbit anti-mouse IgG and donkey anti-mouse IgG. In the case of donkey anti-mouse IgG, each stage of the production of the purified antibody and antibody-enzyme conjugate was monitored by HPLC. HPLC was used to examine monoclonal mouse anti-human T cell and mouse anti-human apoliproprotein B antibodies. The presence of antibody in ascites fluid from mice bearing Landschütz ascites tumour cells was also detected. The antigen-antibody reaction between human serum albumin and anti-human serum albumin was demonstrated using HPLC and this procedure should offer a novel method for studying antigen-antibody interaction.
描述了使用Protein Pak 300 SW柱和Protein Pak DEAE阴离子交换柱通过高效液相色谱（HPLC）对多克隆抗体、单克隆抗体及抗体-酶缀合物进行纯化和表征的方法。检测了以下多克隆抗体：（i）驴抗小鼠免疫球蛋白G（IgG），（ii）小鼠IgG，（iii）羊抗人IgG，以及（iv）山羊抗人因子VIII。HPLC用于分析兔抗小鼠IgG和驴抗小鼠IgG的辣根过氧化物酶缀合物的纯度。对于驴抗小鼠IgG，通过HPLC监测纯化抗体和抗体-酶缀合物生产的每个阶段。HPLC用于检测单克隆小鼠抗人T细胞抗体和小鼠抗人载脂蛋白B抗体。还检测了携带兰施胡茨腹水肿瘤细胞的小鼠腹水中抗体的存在。使用HPLC证明了人血清白蛋白与抗人血清白蛋白之间的抗原-抗体反应，该方法应为研究抗原-抗体相互作用提供一种新方法。
