Automation of Immunoglobulin Glycosylation Analysis

Hendel, Jenifer L.; Gardner, Richard A.; Spencer, Daniel I. R.

doi:10.1007/978-3-030-76912-3_5

Cited by 2 publications

(6 citation statements)

References 47 publications

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“…Studies of this type require a large number of samples to be analysed in a short time, driving the need for the development of high-throughput methodologies for glycosylation analysis ( 14 ). Adopting automated methods would greatly streamline glycome in epidemiology and clinical diagnostics, enhancing the rapid discovery of glycan biomarkers ( 15 ). A study by Groth and Cox found that in 2017, 89% of published life science articles featured a manual protocol with an existing automated alternative ( 16 ).…”

Section: Introductionmentioning

confidence: 99%

“…The digital nature of protocols means effortless transfer between laboratories, making the knowledge more accessible. Lastly, automation eliminates complicated and tedious analysts’ tasks, reduces the risk of exposure to hazardous materials, frees up human resources and lowers costs ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

“…These benefits come with particular challenges. The biggest challenge is the complexity of methods used in glycomics ( 15 ). Pipetting systems, while very precise, come with a set of limitations of their own.…”

Section: Introductionmentioning

confidence: 99%

“…Robots are also challenging to control as they require additional knowledge of instrument software. The last challenge with introducing such systems in laboratories is the exorbitant prices, that make it hard for many research teams to acquire such a device ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

“…Multipurpose liquid-handling robotic workstations have been designed to automate much of the sampling, mixing and manipulating of liquid samples. The two main types of automated liquid-handling workstations used in glycomics laboratories fit into one of the two robotic configurations: Cartesian configuration (moves in three perpendicular axes (X, Y and Z) that allow precise movements and positioning of the robotic arm) and the anthropomorphic or articulated configuration (human-like or arm-like configuration with multiple joints for movement) ( 15 ). There were attempts to develop methods for antibody glycan analysis using Biomek FX manufactured by Beckman Coulter, using MICROLAB STARlet developed by Hamilton, Andrew Alliance semi-automation platform and SweetBlot 7 automated system from System Instruments Co. ( 18 - 23 ).…”

Section: Introductionmentioning

confidence: 99%

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Automated high throughput IgG N-glycosylation sample preparation method development on the Tecan Freedom EVO platform

Lauc,

Novokmet,

Trbojević-Akmačić

et al. 2024

Biochem. med. (Online)

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Graphical abstract Highlights • An automated immunoglobulin G (IgG) N-glycan method was developed leveraging positive pressure technology • In general, automated method showed satisfactory precision and agreement with manual method that is in current use • The automated method holds the possibility to significantly streamline sample preparation, enhancing throughput and improving laboratory safety IntroductionGlycomics, 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. Materials and methodsThe 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. ResultsThe 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. ConclusionsThe 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.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Automated high throughput IgG N-glycosylation sample preparation method development on the Tecan Freedom EVO platform

Lauc,

Novokmet,

Trbojević-Akmačić

et al. 2024

Biochem. med. (Online)

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

Harvey

2024

Mass Spectrometry Reviews

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The use of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry for the analysis of carbohydrates and glycoconjugates is a well‐established technique and this review is the 12th update of the original article published in 1999 and brings coverage of the literature to the end of 2022. As with previous review, this review also includes a few papers that describe methods appropriate to analysis by MALDI, such as sample preparation, even though the ionization method is not MALDI. The review follows the same format as previous reviews. It is divided into three sections: (1) general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation, quantification and the use of computer software for structural identification. (2) Applications to various structural types such as oligo‐ and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals, and (3) other general areas such as medicine, industrial processes, natural products and glycan synthesis where MALDI is extensively used. Much of the material relating to applications is presented in tabular form. MALDI is still an ideal technique for carbohydrate analysis, particularly in its ability to produce single ions from each analyte and advancements in the technique and range of applications show little sign of diminishing.

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Automation of Immunoglobulin Glycosylation Analysis

Cited by 2 publications

References 47 publications

Automated high throughput IgG N-glycosylation sample preparation method development on the Tecan Freedom EVO platform

Automated high throughput IgG N-glycosylation sample preparation method development on the Tecan Freedom EVO platform

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

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