PDF(Pubmed)
Abstract：
UNASSIGNED: Glycomics, focusing on the role of glycans in biological processes, particularly their influence on the folding, stability and receptor interactions of glycoconjugates like antibodies, is vital for our understanding of biology. Changes in immunoglobulin G (IgG) N-glycosylation have been associated with various physiological and pathophysiological conditions. Nevertheless, time-consuming manual sample preparation is one of the limitations in the glycomics diagnostic implementation. The study aimed to develop an automated method for sample preparation on the Tecan Freedom Evo 200 platform and compare its efficiency and precision with the manual counterpart.
UNASSIGNED: The initial method development included 32 pooled blood plasma technical replicates. An additional 24 pooled samples were used in the method comparison along with 78 random duplicates of plasma samples collected from 10,001 Dalmatians biobank to compare the manual and automated methods.
UNASSIGNED: The development resulted in a new automated method. For the automated method, glycan peaks comprising 91% of the total sample glycan showed a variation of less than 5% while 92% of the total sample showed a variation of less than 5% for the manual method. The results of the Passing-Bablok regression indicated no differences between the automated and manual methods for 12 glycan peaks (GPs). However, for 8 GPs systematic difference was present, while both systematic and proportional differences were present for four GPs.
UNASSIGNED: The developed automated sample preparation method for IgG glycan analysis reduced exposure to hazardous chemicals and offered a simplified workflow. Despite slight differences between the methods, the new automated method showed high precision and proved to be highly comparable to its manual counterpart.
UNASSIGNED: The initial method development included 32 pooled blood plasma technical replicates. An additional 24 pooled samples were used in the method comparison along with 78 random duplicates of plasma samples collected from 10,001 Dalmatians biobank to compare the manual and automated methods.
UNASSIGNED: The development resulted in a new automated method. For the automated method, glycan peaks comprising 91% of the total sample glycan showed a variation of less than 5% while 92% of the total sample showed a variation of less than 5% for the manual method. The results of the Passing-Bablok regression indicated no differences between the automated and manual methods for 12 glycan peaks (GPs). However, for 8 GPs systematic difference was present, while both systematic and proportional differences were present for four GPs.
UNASSIGNED: The developed automated sample preparation method for IgG glycan analysis reduced exposure to hazardous chemicals and offered a simplified workflow. Despite slight differences between the methods, the new automated method showed high precision and proved to be highly comparable to its manual counterpart.
摘要：
■糖组学，关注聚糖在生物过程中的作用，特别是它们对折叠的影响，糖缀合物样抗体的稳定性和受体相互作用，对我们理解生物学至关重要。免疫球蛋白G（IgG）N-糖基化的变化与各种生理和病理生理条件有关。然而，耗时的手动样品制备是糖组学诊断实施中的限制之一。该研究旨在开发一种在TecanFreedomEvo200平台上进行样品制备的自动化方法，并将其效率和精度与手动方法进行比较。
■最初的方法开发包括32个混合血浆技术重复。在方法比较中使用另外24个汇集的样品以及从10,001Dalmatians生物库收集的78个随机重复的血浆样品,以比较手动和自动方法。
■开发产生了一种新的自动化方法。对于自动化方法，对于手动方法,包含91%的总样品聚糖的聚糖峰显示小于5%的变化,而92%的总样品显示小于5%的变化。Passing-Bablok回归的结果表明12个聚糖峰(GP)的自动和手动方法之间没有差异。然而,对于8个GP存在系统差异，而四个全科医生存在系统和比例差异。
■开发的用于IgG聚糖分析的自动化样品制备方法减少了对危险化学品的暴露，并提供了简化的工作流程。尽管方法之间略有不同，新的自动化方法显示出高精度，并被证明与手动方法高度可比。
■最初的方法开发包括32个混合血浆技术重复。在方法比较中使用另外24个汇集的样品以及从10,001Dalmatians生物库收集的78个随机重复的血浆样品,以比较手动和自动方法。
■开发产生了一种新的自动化方法。对于自动化方法，对于手动方法,包含91%的总样品聚糖的聚糖峰显示小于5%的变化,而92%的总样品显示小于5%的变化。Passing-Bablok回归的结果表明12个聚糖峰(GP)的自动和手动方法之间没有差异。然而,对于8个GP存在系统差异，而四个全科医生存在系统和比例差异。
■开发的用于IgG聚糖分析的自动化样品制备方法减少了对危险化学品的暴露，并提供了简化的工作流程。尽管方法之间略有不同，新的自动化方法显示出高精度，并被证明与手动方法高度可比。
