Monica T J, Goochee C F, Maiorella B L
Department of Chemical Engineering, Stanford University, CA 94305.
Biotechnology (N Y). 1993 Apr;11(4):512-5. doi: 10.1038/nbt0493-512.
We have conducted a comparative analysis of a monoclonal human IgM obtained from cells cultured in nude-mouse ascites and from the same cells cultured in a bioreactor. We studied the glycosylation of the IgMs using lectin blotting and high-pH anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD), and we also developed reverse phase liquid chromatography (RPLC) peptide maps of the IgM samples. The HPAE-PAD data indicate that the samples differ in both the type and distribution of oligosaccharides present on the IgMs. In addition, the proteins differ in their solubility behavior and in their RPLC peptide maps. We conclude that the method of cell culture is capable of significantly altering the characteristics of the glycoprotein product.
我们对从裸鼠腹水中培养的细胞以及在生物反应器中培养的相同细胞所获得的单克隆人IgM进行了比较分析。我们使用凝集素印迹法和带脉冲安培检测的高pH阴离子交换色谱法（HPAE-PAD）研究了IgM的糖基化情况，并且还绘制了IgM样品的反相液相色谱（RPLC）肽图。HPAE-PAD数据表明，这些样品在IgM上存在的寡糖类型和分布方面均存在差异。此外，这些蛋白质在其溶解行为和RPLC肽图方面也有所不同。我们得出结论，细胞培养方法能够显著改变糖蛋白产物的特性。
