Development of a microarray-based method for allergen-specific IgE and IgG4 detection

Feyzkhanova, Guzel; Voloshin, S A; Smoldovskaya, Olga; Arefieva, Alla; Filippova, M. A.; Barsky, V. E.; Pavlushkina, Ludmila; Butvilovskaya, V. I.; Tikhonov, A. A.; Reznikov, Yuri; Rubina, A. Yu.

doi:10.1186/s12014-016-9136-7

Cited by 26 publications

(26 citation statements)

References 21 publications

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“…As shown in Figure 4a, four major sialylated O-glycans were identified after pT modification, including NS, NG, N 2 S and N 2 G, which are listed in order of decreasing relative abundance. These results are consistent with findings reported in the literature 17 , and a similar protocol has recently been applied for analysis of glycosylation in ovarian cancer cells 60 .…”

Section: Anticipated Resultssupporting

confidence: 92%

“…A total of 80 O-GalNAc glycans were identified from MPS3. The identified O-glycans are consistent with what has been reported in the literature 60 . Image adapted with permission from ref.…”

Section: Figuresupporting

confidence: 91%

“…( a ) Flowchart of the protocol, including references to the steps in the PROCEDURE. ( b ) Chemoenzymatic method for modification and release of glycans: immobilization of glycoproteins on GIG resin by reductive amination; denaturing of glycoproteins using buffer (0.5% SDS, 50 mM DTT) at 100°C for 10 min; modification of sialic acid residues by carbodiimide coupling by 1 M pT in 1 M HCl in the presence of EDC (pH 4-6) at room temperature for 3 h; enzymatic release of N-glycans using 2 μl of PNGase F and 40 μl of GlycoBuffer (10×, 500 mM sodium phosphate) in 358 μl of DI water, followed by chemical release of O-glycans by treatment of GIG resin with 200 μl of ammonium hydroxide (NH 4 OH; 26-28%) and 500 mM PMP in 300 μl of methanol and incubation at 55°C for 24 h. The released O-glycans are purified using a C18 cartridge, and N-glycans are purified using a Carbograph SPE column 20,37,60 . ( c ) Molecular structure of QUANTITY, consisting of a reporter, a balancer and a reactive group.…”

Section: Figurementioning

confidence: 99%

“…Image adapted with permission from ref. 60, BioMed Central (open-access license: https://creativecommons.org/licenses/by/4.0/). …”

Section: Figurementioning

confidence: 99%

“…(i) N-glycans are released from the solid phase by PNGase F. (ii) O-glycans are cleaved using ammonium hydroxide (mild β-elimination) in the presence of 0.5 M PMP (1-phenyl-3-methyl-5-pyrazolone). (iii) Any remaining N-glycans are released by further PNGase F digestion 60 . Image adapted with permission from ref.…”

Section: Figurementioning

confidence: 99%

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Simultaneous quantification of N- and O-glycans using a solid-phase method

Yang

Sokoll

et al. 2017

Nat Protoc

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Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. N-glycans can be released by glycosidases, whereas O-glycans are often cleaved by chemical reaction. However, it is challenging to combine these enzymatic and chemical reactions in order to analyze both N- and O-glycans. We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)–MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. The GIG protocol takes ~3 d.

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Section: Anticipated Resultssupporting

confidence: 92%

Section: Figuresupporting

confidence: 91%

Section: Figurementioning

confidence: 99%

“…Image adapted with permission from ref. 60, BioMed Central (open-access license: https://creativecommons.org/licenses/by/4.0/). …”

Section: Figurementioning

confidence: 99%

Section: Figurementioning

confidence: 99%

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Simultaneous quantification of N- and O-glycans using a solid-phase method

Yang

Sokoll

et al. 2017

Nat Protoc

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Performance evaluation of a laboratory‐developed light‐initiated chemiluminescence assay for quantification of egg white‐specific IgE

Tan

Zhang

Zheng

et al. 2022

Clinical Laboratory Analysis

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Background: Specific IgE (sIgE) testing has become one of the most important tools for diagnosing IgE-mediated food allergy. Enzyme-linked immunosorbent assay (ELISA) and dot-enzyme-linked immunosorbent assay (Dot-ELISA) have been used to measure sIgE in clinical widely. Light-initiated chemiluminescence assay (LICA) is a new method for measuring allergen-sIgE. We aimed to establish a LICA method for quantitative detection of egg white-sIgE and evaluate its performances. Methods:The best chemibeads coupling method in detecting egg white-sIgE was selected, and a LICA method for quantitative detection of egg white-sIgE was established. The precision study was performed according to Clinical and Laboratory Standards Institute (CLSI) EP5-A2. Detection capability which contains limit of blank (LoB), limit of detection (LoD), and limit of quantitation (LoQ) was evaluated according to National Health Commission of the People's Republic of China (NHC) WS/T 514-2017. Linear range was evaluated according to CLSI EP6-A. All data were analyzed using SPSS software.Results: Precision contains repeatability and intermediate precision. The CV of repeatability ranged from 2.72% to 7.29%, and the CV of intermediate precision ranged from 4.93% to 8.64%. The LoB, LoD, and LoQ of the assay were 0.000 kUA/L, 0.053 kUA/L, and 0.076 kUA/L. The assay linear range was 0.076-34.125 kU A /L (r = 0.9979 ≥ 0.9900). Conclusion:This laboratory-developed LICA method can detect egg white-sIgE, and performance meets clinical requirements. This method shows rapid turnaround cycles and high sensitivity. It can be used as an alternative method for clinical detection of egg white-sIgE.

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Well-Based Multiplex Food Allergen Colorimetric ELISA

Matson

2023

Methods in Molecular Biology

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Development of a microarray-based method for allergen-specific IgE and IgG4 detection

Cited by 26 publications

References 21 publications

Simultaneous quantification of N- and O-glycans using a solid-phase method

Simultaneous quantification of N- and O-glycans using a solid-phase method

Performance evaluation of a laboratory‐developed light‐initiated chemiluminescence assay for quantification of egg white‐specific IgE

Well-Based Multiplex Food Allergen Colorimetric ELISA

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