以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758...以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758μg cm^(2),当pH值为5.5~8.0、无机盐浓度为0.3%~1.5%时,rhEPO的检测回收率基本不受影响。对质量浓度为6~15 ng mL的rhEPO水溶液进行检测时,rhEPO检测回收率的平均值为81.9%;对质量浓度为5~20 ng mL的rhEPO人体尿液进行检测时,rhEPO检测回收率的平均值为79.8%。展开更多
Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is a rather new ’soft ionization’ techniques. Because of its high sensitivity and accuracy, it has been widely used in dete...Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is a rather new ’soft ionization’ techniques. Because of its high sensitivity and accuracy, it has been widely used in detection and characterization of macromolecules such as peptides, proteins and olignucleotides. It also has been applied successfully in the analysis of posttranslational modification of proteins, such as phosphorylation and glycosylation. Compared with the conventional chemical methods, mass spectrometry is simple, rapid and does not require radiolabeling. In this research, a systematic method to analyse glycoprotein by MALDI-TOF-MS was developed. A practical sample-recombinant human erythropoietin was analysed by the method. The molecular weight, content of carbohydrate and glycosylation sites of the glycoprotein were determined by MALDI-TOF-MS, combined with the endo-glycosidase digestion and peptide mapping. The experimental result shows that MALDI-TOF-MS is a very powerful technique in the characterization of glycosylation proteins.展开更多
文摘以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758μg cm^(2),当pH值为5.5~8.0、无机盐浓度为0.3%~1.5%时,rhEPO的检测回收率基本不受影响。对质量浓度为6~15 ng mL的rhEPO水溶液进行检测时,rhEPO检测回收率的平均值为81.9%;对质量浓度为5~20 ng mL的rhEPO人体尿液进行检测时,rhEPO检测回收率的平均值为79.8%。
文摘Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is a rather new ’soft ionization’ techniques. Because of its high sensitivity and accuracy, it has been widely used in detection and characterization of macromolecules such as peptides, proteins and olignucleotides. It also has been applied successfully in the analysis of posttranslational modification of proteins, such as phosphorylation and glycosylation. Compared with the conventional chemical methods, mass spectrometry is simple, rapid and does not require radiolabeling. In this research, a systematic method to analyse glycoprotein by MALDI-TOF-MS was developed. A practical sample-recombinant human erythropoietin was analysed by the method. The molecular weight, content of carbohydrate and glycosylation sites of the glycoprotein were determined by MALDI-TOF-MS, combined with the endo-glycosidase digestion and peptide mapping. The experimental result shows that MALDI-TOF-MS is a very powerful technique in the characterization of glycosylation proteins.
