Peng J H, Sheng L S, Xiang B R, An D K
Analytical and Computer Center, China Pharmaceutical University, Nanjing 210009, China.
Yao Xue Xue Bao. 2000 Oct;35(10):764-9.
To elucidate the microheterogeneity of three N-linked oligosaccharide sites of the Chinese-made recombinant human erythropoietin (rHuEPO).
Glu-C digestion, RP-HPLC separation, online HPLC/electrospray ionization mass spectrometry and matrix-assisted laser desorption/ionization time of flight mass spectrometry.
The sialic acid was analyzed directly. Almost every oligosaccharide was acetylated, the acetylation of tetraantennary + 2LacNAc + 4SA and tetraantennary + 2LacNAc + 4SA were reported.
The acetylation of multi-antennary oligosaccharide will improve the activity of rHuEPO in vivo. The biantennary oligosaccharide was found mainly existing at N-24. For the first time, the carbohydrate structures of each N-linked glycosylated site of Chinese-made rHuEPO were reported.
阐明国产重组人促红细胞生成素(rHuEPO)三个N-连接寡糖位点的微观异质性。
Glu-C酶切、反相高效液相色谱分离、在线高效液相色谱/电喷雾电离质谱和基质辅助激光解吸/电离飞行时间质谱。
直接分析唾液酸。几乎每个寡糖都被乙酰化,报道了四天线 + 2乳糖胺 + 4唾液酸和四天线 + 2乳糖胺 + 4唾液酸的乙酰化情况。
多天线寡糖的乙酰化将提高rHuEPO在体内的活性。发现双天线寡糖主要存在于N-24位。首次报道了国产rHuEPO每个N-连接糖基化位点的碳水化合物结构。