质谱法:用于碳水化合物-蛋白质偶联监测和糖缀合物分子质量测定的强大工具。

IF 2.7 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Glycoconjugate Journal Pub Date : 2023-08-01 DOI:10.1007/s10719-023-10126-7
Di Wu, Peng Xu, Meagan Kelly, Edward T Ryan, Pavol Kováč, Grzegorz Piszczek
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引用次数: 0

摘要

糖缀合疫苗是预防细菌和病毒病原体引起的疾病的现有手段的重要补充。将碳水化合物与蛋白质结合是开发这些疫苗的关键一步。传统的质谱技术,如MALDI-TOF和SELDI-TOF,在检测高分子质量的糖缀合物方面存在困难。质谱测光(MP)是最近发展起来的一种单分子技术,它可以测量单个分子的质量,并根据数百到数千个这些测量结果生成质量分布。在这项研究中,我们评估了MP在监测碳水化合物-蛋白质偶联反应和表征偶联物方面的性能。用载体蛋白BSA制备了三种不同的糖缀合物,另一种糖缀合物来自一个分子量为3.74 MDa的病毒衣壳蛋白复合体。MP测定的质量与SELDI-TOF-MS和SEC-MALS测定的质量一致。牛血清白蛋白二聚体与碳水化合物抗原的结合也被成功表征。这项研究表明,MP技术是早期监测糖缀合反应和表征糖缀合物的一种有前途的替代方法。它测量溶液中完整的分子,在很宽的质量范围内精度很高。MP只需要非常少量的样品,并且没有特定的缓冲限制。MP的其他优势包括耗材成本最低,数据收集和分析速度快。与其他方法相比,它的优点使其成为糖缀合领域研究人员的宝贵工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Mass photometry: A powerful tool for carbohydrates-proteins conjugation monitoring and glycoconjugates molecular mass determination.

Glycoconjugate vaccines are important additions to the existing means for prevention of diseases caused by bacterial and viral pathogens. Conjugating carbohydrates to proteins is a crucial step in the development of these vaccines. Traditional mass spectrometry techniques, such as MALDI-TOF and SELDI-TOF, have difficulties in detecting glycoconjugates with high molecular masses. Mass photometry (MP) is a single-molecule technique that has been recently developed, which allows mass measurements of individual molecules and generates mass distributions based on hundreds to thousands of these measurements. In this study, we evaluated the performance of MP in monitoring carbohydrate-protein conjugation reactions and characterization of conjugates. Three different glycoconjugates were prepared from carrier protein BSA, and one from a large protein complex, a virus capsid with 3.74 MDa molecular mass. The masses measured by MP were consistent with those obtained by SELDI-TOF-MS and SEC-MALS. The conjugation of BSA dimer to carbohydrate antigen was also successfully characterized. This study shows that the MP technique is a promising alternative to methods developed earlier for monitoring glycoconjugation reactions and characterization of glycoconjugates. It measures intact molecules in solution and it is highly accurate over a wide mass range. MP requires only a very small amount of sample and has no specific buffer constraints. Other MP advantages include minimal cost of consumables and rapid data collection and analysis. Its advantages over other methods make it a valuable tool for researchers in the glycoconjugation field.

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来源期刊
Glycoconjugate Journal
Glycoconjugate Journal 生物-生化与分子生物学
CiteScore
6.00
自引率
3.30%
发文量
63
审稿时长
1 months
期刊介绍: Glycoconjugate Journal publishes articles and reviews on all areas concerned with: function, composition, structure, biosynthesis, degradation, interactions, recognition and chemo-enzymatic synthesis of glycoconjugates (glycoproteins, glycolipids, oligosaccharides, polysaccharides and proteoglycans), biochemistry, molecular biology, biotechnology, immunology and cell biology of glycoconjugates, aspects related to disease processes (immunological, inflammatory, arthritic infections, metabolic disorders, malignancy, neurological disorders), structural and functional glycomics, glycoimmunology, glycovaccines, organic synthesis of glycoconjugates and the development of methodologies if biologically relevant, glycosylation changes in disease if focused on either the discovery of a novel disease marker or the improved understanding of some basic pathological mechanism, articles on the effects of toxicological agents (alcohol, tobacco, narcotics, environmental agents) on glycosylation, and the use of glycotherapeutics. Glycoconjugate Journal is the official journal of the International Glycoconjugate Organization, which is responsible for organizing the biennial International Symposia on Glycoconjugates.
期刊最新文献
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