Lectin Bead Array in a Single Tip Facilitates Fully Automatic Glycoprotein Profiling

Shimazaki, Hiroko; Saito, Kozue; Matsuda, Atsushi; Sawakami, Kazumi; Kariya, Minoru; Segawa, Osamu; Miyashita, Yukiko; Ueda, Tetsuya; Koizuka, Michinori; Nakamura, Kazuhiro; Kaji, Hiroyuki; Tajima, Hideji; Kuno, Atsushi

doi:10.1021/acs.analchem.9b01876

Cited by 10 publications

(4 citation statements)

References 35 publications

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“…However, lectins are able to exist in oligomer structures with multiple binding sites to simultaneously interact with different glycans of the same glycoprotein or with different arms of a branched oligosaccharide, thus achieving multivalent interaction to boost selectivity and affinity . Lectins have been utilized as capable recognition element to enable glycoprotein identification, profiling and sensing. − Tkac and co-workers have developed a series of lectin-based impedimetric biosensors for the sensitive detection and glycoprofiling of glycoproteins. , Recently, Qu and co-worker reported to modify lectin of concanavalin A (Con A) on metal–organic framework material of MIL-101(Cr)-NH 2 to afford an electrochemical impedimetric biosensing platform of Con A@MIL-101(Cr)-NH 2 for glycoprotein detection …”

Section: Receptors For Glycoproteinsmentioning

confidence: 99%

Functional Interface for Glycoprotein Sensing: Focusing on Biosensors

Qu,

2024

Langmuir

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Glycosylated proteins or glycoproteins make up a large family of glycoconjugates, and they participate in a variety of fundamental biological events. Glycoproteins have become important biomarkers in the diagnosis and treatment of a number of tumors. Biosensors are quite suitable for glycoprotein detection. The design and fabrication of a functional sensing interface play a crucial role in the biosensor construction to target glycoproteins. The functional interface, particularly receptors, typically determines the key characteristics of a biosensor, such as selectivity and sensitivity. Antibody, peptide, aptamer, boronic acid derivative, lectin, and molecularly imprinted polymer are all capable receptors for glycoprotein recognition, and each of these will be discussed. Most glycoproteins exist in low abundance, thus rendering signal amplification techniques indispensable. Nucleic acid-mediated and nanomaterial-mediated signal amplification for the detection of glycoproteins will be focused on herein. This review aims to highlight these different functional interfaces for glycoprotein sensing.

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Section: Receptors For Glycoproteinsmentioning

confidence: 99%

Functional Interface for Glycoprotein Sensing: Focusing on Biosensors

Qu,

2024

Langmuir

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“…By contrast, GlycoBIST performs measurements of around 10‐plex interactions to obtain quick glycan profiling within 2 hr of tip preparation. The adoption of chemiluminescent detection for GlycoBIST allows measurements in a wide dynamic range compared with fluorescence‐based lectin microarray technology (Shimazaki et al., 2019). In addition, the 10‐plex level of the customized lectin array combined with the preliminary selection afforded by the 45‐plex lectin microarray is suitable in large‐scale experiments for the following two reasons: (1) Up to 1000 probe (lectin) beads are potentially prepared at the same time that would be used as one lot of tips.…”

Section: Commentarymentioning

confidence: 99%

“…Among the many analytical methods available for the determination of glycan structure, lectin microarray has been used widely to understand and compare the characteristics of glycan structure without liberation of the glycan (Hirabayashi, Yamada, Kuno, & Tateno, 2013). The protocol introduced here (i.e., lectin bead array in a single tip [GlycoBIST]), is a technique that aims to be a simple and reproducible lectin microarray analysis using an automated pipetting machine and a bead array with a diameter of 1 mm lined up in a tip, enabling glycan profiling to become a routine task without a requirement for specific expertise in glycoscience (Shimazaki et al., 2019).…”

Section: Introductionmentioning

confidence: 99%

GlycoBIST: A System for Automatic Glycan Profiling of a Target Protein Using Milli‐Bead Array in a Tip

et al. 2020

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Lectin is a biomolecule that recognizes a specific part of glycans and, thus, has been used widely as a probe for glycoprotein analysis. Owing to the wide repertoire in nature combined with the recent two decades of advances in microarray technology, the multiplexed use of lectins has been widely used for glycan profiling of endogenous proteins. Because protein glycosylation is recognized as being biologically important and is expected to be a reliable disease marker, lectin microarray analysis with highly sensitive detection has been used to discover disease‐relevant glycosylation alterations. However, the conventional system is limited to research purposes; thus, its implementation in clinical settings is warranted. Here, we provide an automatic glycan profiling method using GlycoBIST. A unique array format is used for 10‐plexed lectin–glycoprotein interaction analysis on 1‐mm‐sized beads, which are arranged vertically in a capillary‐shaped plastic tip. Using a one‐boxed autopipetting machine, the whole process (including interaction, washing, and detection) is performed automatically and serially, resulting in reproducible measurements. In this article, a typical method for glycan profiling of a purified glycoprotein and the fabrication of GlycoBIST tips is explained. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Fabrication of a GlycoBIST tip Basic Protocol 2: Automatic profiling of a target glycoprotein using GlycoBIST

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“…In the first attempt to automate this process, we have previously reported the milli-sized lectin bead array system GlycoBIST, 13 enabling minimum glycan profiling with 10lectins handled by simple robotics, with potential automatic data processing for signal quantification. 14 Notably, it is suitable for a large-scale experiment, as up to 1000 lectin beads for one lectin are prepared simultaneously and maintained in the same product batch before GlycoBIST tip production.…”

mentioning

confidence: 99%

Auto-Lectin Dotcoding by Two Octopuses: Rapid Analysis of Fluorescence-Labeled Glycoproteins by an 8-channel Fully-Automatic Bead Array Scanner with a Rolling-Circle Detector

et al. 2023

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Protein glycosylation is a crucial factor that must be evaluated in biological pharmaceuticals. The glycoform profile of a protein can vary depending on the conditions of the cultivation, purification process, and the selection of a host cell. Lectin microarrays are reliable bioanalytical methods used in the early phases of bioprocesses for the detection of glycosylation. The concept of a fully automated glycan detection with a bead array has been previously reported; however, no simple system has been constructed on fluorescence-based detection using a microarray. Here, we present a fully automated detection system equipped with a novel fluorescence detector for a 13-lectin bead array with a single tip. The lattice-like arrangement of a set of fibers proximate to the tip of the light emitting diode and photomultiplier tube detector minimized the noise caused by the reflection of incident light on the plastic capillary tip and bead. A unique rolling-circle fiber unit with quadruple lattices stacked in two layers realizes the 8-parallel automeasurement with a drastic reduction in scanning time and machine size. The 8-glycan profiles obtained automatically within 25 min were identical with those obtained with the conventional lectin microarray after overnight incubation. The signals obtained were represented as lectin dotcodes. Therefore, autolectin dotcoding assisted by the twin 8 legs named as “detection and irradiation octopuses” may be a rapid glyco-evaluation system during the production and development of biopharmaceuticals.

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Lectin Bead Array in a Single Tip Facilitates Fully Automatic Glycoprotein Profiling

Cited by 10 publications

References 35 publications

Functional Interface for Glycoprotein Sensing: Focusing on Biosensors

Functional Interface for Glycoprotein Sensing: Focusing on Biosensors

GlycoBIST: A System for Automatic Glycan Profiling of a Target Protein Using Milli‐Bead Array in a Tip

Auto-Lectin Dotcoding by Two Octopuses: Rapid Analysis of Fluorescence-Labeled Glycoproteins by an 8-channel Fully-Automatic Bead Array Scanner with a Rolling-Circle Detector

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