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目的建立一种更加灵敏高效的糖蛋白染色鉴别方法。方法在高碘酸-Schiff法的基础上,通过对固定、还原、氧化等溶液配方及染色时间的改变,对电泳凝胶糖蛋白染色法进行改进,并与传统高碘酸-Schiff法进行比较。结果重组单抗样品经改进的糖蛋白染色法染色,仅重链显色,轻链不显色;再经考马斯亮蓝染液复染,轻、重链均显色;改进的染色方法的糖含量检测灵敏度为0.15μg糖,而传统高碘酸-Schiff法的糖含量检测灵敏度为3μg糖。结论已成功建立了一种新的特异性好、灵敏度高且用时短的糖蛋白染色鉴别方法。
Objective To establish a more sensitive and efficient glycoprotein staining method. Methods Based on periodic acid-Schiff method, the gel electrophoresis method was improved by the method of fixation, reduction and oxidation, and compared with the traditional periodic acid-Schiff method . Results The recombinant mAb was stained by modified glycoprotein stain, only the heavy chain color was developed and the light chain did not develop color. After reconstitution with Coomassie brilliant blue dye, the light and heavy chains were both developed. Improved staining method of sugar The detection sensitivity of 0.15μg sugar, while the traditional periodic acid - Schiff method of sugar content detection sensitivity of 3μg sugar. Conclusion A new method of identifying glycoprotein staining with high specificity, high sensitivity and short duration has been successfully established.