We describe a simplified approach for purification and characterization of human ‘IgG-Fc’ fragment used widely as immunochemical tool and for therapeutic purposes. The ‘Fc’ fragment was purified from human IgG in a 3-stage column chromatography. The purified ‘Fc’ fragment appeared as a dimer glycoprotein with an apparent molecular mass of 52981 Dalton (Ultraflex MALDI TOF/TOF). The Size-exclusion HPLC profile of the purified ‘Fc’ fragment of human IgG matched that of a commercially procured reference ‘Fc’ fragment material. The purity of the ‘Fc’ fragments was >99% by SDS-PAGE and size-exclusion HPLC. The results of Western blotting, immunoelectrophoresis, and mass spectrometry analysis indicate a high purity of the ‘Fc’ fragment. Peptide mass fingerprint analysis of the purified ‘Fc’ protein yielded peptides that partially match the known database sequences of FCG3B_HUMAN (Uniprot ID: O75015). This method of purification of the ‘Fc’ fragment is suitable for achieving high purity level of ‘Fc’ fragment protein. With this purification approach, the cost of the purified ‘Fc’ fragment of human IgG is significantly reduced as compared with the current market price of IgG-Fc fragment protein in international market. The purified ‘IgG-Fc’ fragment protein was found to be negative for major viral markers.

我们描述了纯化和表征人的“ IgG-Fc”片段的简化方法，该片段广泛用作免疫化学工具和用于治疗目的。在三阶段柱色谱法中从人IgG中纯化“ Fc”片段。纯化的“ Fc”片段显示为二聚体糖蛋白，表观分子量为52981道尔顿（Ultraflex MALDI TOF / TOF）。纯化的人IgG'Fc'片段的大小排阻HPLC谱与市售参比'Fc'片段物质的HPLC相匹配。通过SDS-PAGE和尺寸排阻HPLC，“ Fc”片段的纯度＞ 99％。Western印迹，免疫电泳和质谱分析的结果表明“ Fc”片段的纯度很高。纯化的“ Fc”蛋白的肽质量指纹分析得到的肽与FCG3B_HUMAN（Uniprot ID：O75015）的已知数据库序列部分匹配。纯化“ Fc”片段的这种方法适合于实现“ Fc”片段蛋白的高纯度水平。通过这种纯化方法，与国际市场上IgG-Fc片段蛋白的当前市场价格相比，纯化的人IgG'Fc'片段的成本大大降低。发现纯化的“ IgG-Fc”片段蛋白对主要病毒标志物呈阴性。与国际市场上IgG-Fc片段蛋白的当前市场价格相比，纯化的人IgG'Fc'片段的成本大大降低。发现纯化的“ IgG-Fc”片段蛋白对主要病毒标志物呈阴性。与国际市场上IgG-Fc片段蛋白的当前市场价格相比，纯化的人IgG'Fc'片段的成本大大降低。发现纯化的“ IgG-Fc”片段蛋白对主要病毒标志物呈阴性。