Comparison of Methods of Immobilization to Enzyme-Linked Immunosorbent Assay Plates for the Detection of Sugar Chains

Satoh, Ayano; Fukui, Emiko; Yoshino, Saori; Shinoda, Mayumi; Kojima, Kensuke; Matsumoto, Isamu

doi:10.1006/abio.1999.4329

Cited by 16 publications

(11 citation statements)

References 10 publications

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“…Saccharides were attached by 1-OH to reactive groups of this copolymer using bifunctional spacer containing hydrazide or amino functions. Despite the simplicity of the method, not all saccharides were immobilized successfully with the copolymer and not all of them displayed biological activity after immobilization [17,18]. 1-OH derivatives of mono-and simple oligosaccharides were covalently linked to the solid surface via long hydrophobic cyanurchloride-activated linker [19].…”

Section: Discussionmentioning

confidence: 99%

High molecular weight neoglycoconjugates for solid phase assays

et al. 2005

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Adsorption of a carbohydrate on solid phase is the necessary stage of the immunosorbent assay (ELISA) and analogous methods of the study of carbohydrate-protein interaction. Usually physical adsorption on polystyrene requires a high concentration of conjugated carbohydrate and, thus, enormous consumption of it. In this study, we explored two approaches allowing more rational use of oligosaccharide (Glyc). The first of them is based on the covalent immobilization of neoglycoconjugates on the NH(2)-modified polystyrene; the second one is based on the elevated adherence of high m.w. neoglycoconjugates to polystyrene. Covalent immobilization of polyacrylamide conjugates, Glyc-PAA, provided a possibility to solve the problem, but the nonspecific binding of antibodies in ELISA proved to be unacceptably high. At the same time, the increase of the Glyc-PAA m.w. from 30 kDa to 2,000 kDa allowed a 10-20 fold decrease of its consumption, when using physical adsorption, whereas the assay background remained at the low level. The amount of 2,000 kDa Glyc-PAA that is sufficient for the coating of a standard 96-well plate corresponds to the nanomole level of oligosaccharide, this providing a possibility to use saccharides that are available in a very limited amount when studying the carbohydrate-protein interaction with solid-phase techniques.

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Section: Discussionmentioning

confidence: 99%

High molecular weight neoglycoconjugates for solid phase assays

et al. 2005

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“…[28][29][30][31] The main advantages of an oxime linker strategy for solid-phase immobilization of carbohydrates include: (i) ease of formation under mild aqueous conditions and high nucleophilicity via the a-effect; (ii) good stability under a wide range of pH; (iii) versatility in the choice of supports and saccharides. In an ELISA-microtiter plate format, lactose and N-acetylchitooligosaccharides have been immobilized through the reducing end via oxime formation.…”

Section: Methods For the Immobilization Of Unmodified Oligosaccharidesmentioning

confidence: 99%

“…In an ELISA-microtiter plate format, lactose and N-acetylchitooligosaccharides have been immobilized through the reducing end via oxime formation. 29 The aminooxy anchor was installed on the amino-coated plate using Boc-Aoa-OH and N-ethyl-N 0 -(dimethylaminopropyl)carbodiimide as the coupling reagent.…”

Section: Methods For the Immobilization Of Unmodified Oligosaccharidesmentioning

confidence: 99%

“…20 Oligosaccharides immobilized on microtiter (ELISA) plates have been detected by a horseradish peroxidase (HPR)-based colorimetric assay. 29 First, the oligosaccharides were bound to biotinylated or HPR-conjugated lectins in Tris-buffer saline (TBS, pH 7.5). Diluted HPRconjugated biotin-streptavidin complex with TBS was then added.…”

Section: Methods For Determination Of Immobilization Efficiencymentioning

confidence: 99%

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Solid-phase chemical tools for glycobiology

Larsen¹,

Thygesen²,

Guillaumie

et al. 2006

Carbohydrate Research

136

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“…These techniques are often referred to as enzymelinked-lectin assays or ELLA [47]. A variety of linkage techniques have been applied to immobilize the oligosaccharides to the plates [48][49][50] and certain modifications, such as the use of immobilized glycoproteins, have been tested [51]. However, many ELLA techniques have been developed with a focus on determining the properties of the bound carbohydrate residue rather than the quantification of the lectin.…”

Section: Introductionmentioning

confidence: 99%

Validation of a Method for Quantitation of Soybean Lectin in Commercial Varieties

Breeze

Leyva-Guerrero

Yeaman

et al. 2015

J Americ Oil Chem Soc

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for a capture phase and the biotinylated version (Gal-NAc-PAA-Biotin) for detection. Based upon the validation data, the ELLA method can precisely and accurately determine soybean lectin levels in soybean seed. The validated ELLA method was used to quantify SBA in nine commercial soybean varieties introduced between 1972 and 2008 and demonstrated that the natural variability of SBA is subject to the effects of genotype and environment.

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Comparison of Methods of Immobilization to Enzyme-Linked Immunosorbent Assay Plates for the Detection of Sugar Chains

Cited by 16 publications

References 10 publications

High molecular weight neoglycoconjugates for solid phase assays

High molecular weight neoglycoconjugates for solid phase assays

Solid-phase chemical tools for glycobiology

Validation of a Method for Quantitation of Soybean Lectin in Commercial Varieties

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