The Biotin/Avidin-Mediated Microtiter Plate Lectin Assay with the Use of Chemically Modified Glycoprotein Ligand

Duk, Maria; Lisowska, Elwira; Wu, Jun; Wu, Albert M.

doi:10.1006/abio.1994.1410

Cited by 146 publications

(142 citation statements)

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“…It could also be performed on the lower grades of BSA to further reduce costs if necessary. Some protein precipitation was observed during incubation of BSA with the sodium periodate but as BSA solutions were diluted prior to use, this did not affect the reproducibility of the assay We also evaluated methods that reported the omission of blocking steps (26). These methods simply include Tween 20 in lectin probing solutions.…”

Section: Discussionmentioning

confidence: 99%

“…Duk et al reported an ELLA protocol whereby the wells of ELISA plates were coated with glycoprotein (or a glycoconjugate) solutions and biotinylated lectins were added directly to the wells without a prior blocking step (26). They reported that once Tween-20 was included in the lectin solution, absorbance readings in lectin control wells, with no immobilized glycoprotein, was low.…”

Section: Evaluation Of Ella Protocols Omitting Plate Surface Blockingmentioning

confidence: 99%

“…This assay has the same basic format as a standard ELISA (see Figure 1) and allows the analysis of glycoproteins, and lectin-glycan interactions, in a standard microtitre plate format. The most common format of the ELLA requires the immobilization of glycoconjugates on the surface of an ELISA plate, subsequent blocking of the plate surface and then probing with biotinylated lectins (26). The major problem that is encountered when performing the ELLA stems from the lack of suitable blocking reagents.…”

Section: Introductionmentioning

confidence: 99%

“…The major problem that is encountered when performing the ELLA stems from the lack of suitable blocking reagents. Many of the blocking reagents traditionally used in ELISA are unsuitable for use in ELLA as they generate high background signals, contributing to false-positive signals, due to the presence of carbohydrate in the preparations (26)(27)(28)(29). As a result of these difficulties many of the ELLA protocols reported in the literature to date have only utilized small and specific subsets of lectins (30)(31)(32) and their utility is thereby restricted.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis

Thompson

Creavin

O’Connell

et al. 2011

Analytical Biochemistry

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Lectin's are proteins capable of recognising and binding to specific oligosaccharide structures found on glycoproteins and other biomoloecules. As such they have found utility for glycoanalytical applications. One common difficulty encountered in the application of these proteins, particularly in multi-well plate assay formats known as Enzyme Linked Lectin Assays (ELLA's), is in finding appropriate blocking solutions to prevent non-specific binding with plate surfaces. Many commonly used blocking agents contain carbohydrates and generate significant background signals in ELLA's, limiting the utility of the assay.In this study we examined the suitability of a range of blocking reagents, including protein based, synthetic and commercially available carbohydrate free blocking reagents, for ELLA applications. Each blocking reagent was assessed against a panel of 19 commercially available biotinylated lectins exhibiting diverse structures and carbohydrate specificities. We identified the synthetic polymer Polyvinyl Alcohol (PVA) as the best global blocking agent for performing ELLA's. We ultimately present an ELLA methodology facilitating broad spectrum lectin analysis of glycoconjugates and extending the utility of the ELLA.

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Section: Discussionmentioning

confidence: 99%

Section: Evaluation Of Ella Protocols Omitting Plate Surface Blockingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Optimization of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis

Thompson

Creavin

O’Connell

et al. 2011

Analytical Biochemistry

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“…ELLA is based on binding of biotinyled lectins to glycoconjugates immobilized on a microtiter plate. The bound lectins are detected with labeled avidin (streptavidin) (Duk et al 1994;Orntoft et al 1997;Kratz et al 2003). This experimental approach was used to characterize T-antigen levels (Reddi et al 2000).…”

Section: Enzyme-linked Lectin Assays (Ella)mentioning

confidence: 99%

Lectinomics I. Relevance of exogenous plant lectins in biomedical diagnostics

et al. 2009

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This review focuses on utilization of plant lectins as medical diagnostic reagents and tools. The lectin-related diagnostic is aimed at detection of several diseases connected to alteration of the glycosylation profiles of cells and at identification of microbial and viral agents in clinical microbiology. Certain lectins, proposed for or used as diagnostic tools could even recognize those cellular determinants, which are not detected by available antibodies. Broad information is presented on the lectinomics field, illustrating that lectin diagnostics might become practical alternative to antibody-based diagnostic products. In addition, the rising trend of lectin utilization in biomedical diagnostics might initiate a development of innovative methods based on better analytical technologies. Lectin microarray, a rapid and simple methodology, can be viewed as an example for such initiative. This technology could provide simple and efficient screening tools for analysis of glycosylation patterns in biological samples (cellular extracts, tissues and the whole cells), allowing thus personalized detection of changes associated with carbohydrate-related diseases.

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Adhesion of human uroepithelial cells to E-selectin: Possible involvement of sialosyl LewisA-ganglioside

et al. 1996

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In a previous study we showed that tumorigenic and invasive human uroepithelial cell lines are characterized by the presence of sialosyl le' ($Lee') ganglioside. Our data suggested that expression of this glycolipid correlated with acquisition of the malignant phenotype by human urothelial cells. To evaluate the postulated adhesion function of $Lea antigen, we studied the adherence of 6 human urothelial cell lines with different expressions of this carbohydrate structure to E-selectinexpressing CHO cells. The only cell line that bound specifically to E-selectin was Hu 1703He. which expressed the highest level of sl& antigen. The involvement of carbohydrat+E-selectin interaction in the adhesion of Hu 1703He cells was indicated by the following facts: (i) anti-E-selectin monoclonal antibody (MAb) completely abolished binding to E-selectin-expressing CHO cells; (ii) removal of sialic acid from Hu 1703He cells highly decreased the adhesion. Adhesion correlated with the presence of several sLea-carrying glycoproteins, which was shown by immunoblotting of Hu 1703He cell lysate with anti-sle. MAb 19-9. The binding of antibody was abolished when cell lysate was treated with 0-sialoglycoprotein endopeptidase. suggesting that sLea is present on 0-linked oligosaccharides. However, incubation of Hu I703He cells with 0-sialogtycoprotease had no effect on adhesion to E-selectin or on binding of 19-9 MAb to the cell surface. Our data suggest that (I) protein-bound sLea oligosaccharides represent only a minor portion of whole s h e ' anti en roduced by uroepithelial cells; (ii) effective binding to E-sefecti occurs when s h . oligosaccharide present on cellsurface glycorphingolipids is expressed in high densitysince the cell lines with moderate expression of sle' ganglioside did not bind to E-selectin-tramfected CHO cells. 0 1996 Wiley-Lks, lnc. whom correspondence and reprint requests should be sent, at Department of Immunochemistry, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Czerska 12, 53-114 Wroclaw. Poland. Fax: 48-71-67 91 11. Abbmiotions: sLea, sialosyl Lea, sLex, sialosyl Lex, PBS, phos hate buffered saline: TBS, tris-buffered saline; PMSF, phenylmethyfsulfo: nyl fluoride: SDS-PAGE, sodium dodecyl olyacrylamide el electrophoresis; BSA, bovine serum albumin; J A b , monoclonaf antibody; FITC, fluorescein-isothiocyanate KLOPOCKI ETAL.

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The Biotin/Avidin-Mediated Microtiter Plate Lectin Assay with the Use of Chemically Modified Glycoprotein Ligand

Cited by 146 publications

References 0 publications

Optimization of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis

Optimization of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis

Lectinomics I. Relevance of exogenous plant lectins in biomedical diagnostics

Adhesion of human uroepithelial cells to E-selectin: Possible involvement of sialosyl LewisA-ganglioside

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