STRUCTRUAL ANALYSIS OF N-LINKED OLIGOSACCHARIDES IN MEN2234 GLYCOPROTEIN

MEN2234 糖蛋白中 N-连接寡糖的结构分析

• 批准号: 7602836
• 负责人: Parastoo Azadi
• 金额: $ 0.12万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602836
• 关键词: Acetic Acid; Acetic Acids; Acids; Amino Sugars; CD7 gene; Calibration; Computer Retrieval of Information on Scientific Projects Database; Data; Funding; Glycoproteins; Grant; Hydrolysis; Ice; Injection of therapeutic agent; Institution; Link; Monosaccharides; Needles; Oligosaccharides; Pump; Research; Research Personnel; Resources; Running; Sampling; Sialic Acids; Sodium Acetate; Source; Stainless Steel; Standards of Weights and Measures; System; Trifluoroacetic Acid; United States National Institutes of Health; Water; base; detector; prevent; programs

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Neutral and amino sugars were released from 150 ¿L of each of the samples submitted through hydrolysis with 2 N trifluoroacetic acid at 100¿C for 4 h. After hydrolysis the samples were lyophilized. The dried samples were then resuspended in 150 ¿L water and sonicated on ice. For analysis, 10 ¿L were injected. Blanks (water) were injected and run between the standards and the sample (data not shown) to prevent carry-over. The calibration standards were hydrolyzed in parallel with the samples, and then injected at three levels (0.5, 1.0 and 2.0 nmol). The quantitation was based on a linear interpolation from these standards. Sialic acids were released from 150 ¿L of each of the samples submitted by mild acid hydrolysis using 2M acetic acid at 80¿C for 3 h. The samples were dried as above, and resuspended in 150 ¿L water. For analysis, 10 ¿L were injected. Blanks (water) were injected and run between the standards and the sample (data not shown) to prevent carry-over. The calibration standards were hydrolyzed in parallel with your samples, and then injected at three levels (0.5, 1.0 and 2.0 nmol). The quantitation was based on a linear interpolation from these standards. Monosaccharides were analyzed using a Dionex DX500 system equipped with a GP40 gradient pump, an ED40 electrochemical detector and a Thermo-Separations AS3500 autosampler containing a stainless steel needle. Analyses were performed using a Dionex CarboPac PA20 (3 x 150 mm) analytical column with an Amino Trap column. The gradient programs used eluents A, water; B, 100 mM NaOH for the neutral and amino sugars; C, 100 mM NaOH; and D, 1 M sodium acetate in 100 mM NaOH for the sialic acids. Injections were made every 40 minutes for the neutral and amino sugar determinations and every 35 minutes for the sialic acid determinations.

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