New technology for regiospecific covalent coupling of polysaccharide antigens in ELISA for serological detection

Jauho, E. S.; Boas, Ulrik; Wiuff, Camilla; Wredstrøm, K.; Pedersen, Brian; Andresen, Lars Ole; Heegaard, Peter M. H.

doi:10.1016/s0022-1759(00)00248-9

Cited by 18 publications

(11 citation statements)

References 14 publications

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“…However, with day 28 samples (i.e., postvaccination samples with higher responses), the results generated with the bead assay or ELISA were less divergent with the median anti-Vi IgG EU/ml value of 26.1 for the bead assay and 53.1 for ELISA (only a 2-fold difference). Despite other reports indicating that polysaccharide antigens used as ELISA coating antigens provided inconsistent results (3,15,16,26,33), our data suggest that the Vi polysaccharide used as an ELISA coating antigen provided consistent results (as long as the serum contained elevated levels of anti-Vi antibody) and are supported by previous publications that utilize Vi polysaccharide as the coating antigen in ELISA (7,19,21). It seems possible that the purity of the Vi polysaccharide, the type of ELISA reagents utilized (ELISA plates, coating buffers, wash buffers, etc.)…”

Section: Discussionsupporting

confidence: 60%

“…The use of polysaccharides (lipopolysaccharide [LPS], Haemophilus influenzae type b capsular polysaccharide, Vi polysaccharide) as coating antigens for immunoassays is plagued by problems such as a poor binding of polysaccharides to ELISA plates and inconsistent results (3,15,16,26,33). To increase binding of Vi antigen to ELISA plates and produce more-robust assays, others have biotinylated Vi and then added it to streptavidincoated plates (12) or conjugated Vi to tyramine (22,26).…”

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confidence: 99%

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Development of a Bead Immunoassay To Measure Vi Polysaccharide-Specific Serum IgG after Vaccination with the Salmonella enterica Serovar Typhi Vi Polysaccharide

Staats

Kirwan

Whisnant

et al. 2010

Clin Vaccine Immunol

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Vi polysaccharide from Salmonella enterica serotype Typhi is used as one of the available vaccines to prevent typhoid fever. Measurement of Vi-specific serum antibodies after vaccination with Vi polysaccharide by enzyme-linked immunosorbent assay (ELISA) may be complicated due to poor binding of the Vi polysaccharide to ELISA plates resulting in poor reproducibility of measured antibody responses. We chemically conjugated Vi polysaccharide to fluorescent beads and performed studies to determine if a bead-based immunoassay provided a reproducible method to measure vaccine-induced anti-Vi serum IgG antibodies. Compared to ELISA, the Vi bead immunoassay had a lower background and therefore a greater signal-to-noise ratio. The Vi bead immunoassay was used to evaluate serum anti-Vi IgG in 996 subjects from the city of Kolkata, India, before and after vaccination. Due to the location being one where Salmonella serotype Typhi is endemic, approximately 45% of the subjects had protective levels of anti-Vi serum IgG (i.e., 1 g/ml anti-Vi IgG) before vaccination, and nearly 98% of the subjects had protective levels of anti-Vi serum IgG after vaccination. Our results demonstrate that a bead-based immunoassay provides an effective, reproducible method to measure serum anti-Vi IgG responses before and after vaccination with the Vi polysaccharide vaccine.

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Section: Discussionsupporting

confidence: 60%

mentioning

confidence: 99%

Development of a Bead Immunoassay To Measure Vi Polysaccharide-Specific Serum IgG after Vaccination with the Salmonella enterica Serovar Typhi Vi Polysaccharide

Staats

Kirwan

Whisnant

et al. 2010

Clin Vaccine Immunol

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“…Moreover, quite a large amount of saccharide (∼10 nmol/well) is necessary due to low yield of conjugation. Chemical immobilization of polysaccharides on polystyrene using photoactivated anthraquinone attached to 1-OH of polysaccharide has been described in [16]. It has also been proposed to precoat polystyrene with copolymer of methylvinyl ester and maleinic anhydride.…”

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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“…The complete O-antigen polysaccharide (including a portion of the core oligosaccharide) obtained from S. enterica sv. Typhimurium LPS by mild acid hydrolysis [23], was also conjugated similarly. (Fig.…”

Section: Preparation Of Salmonella O-antigen Microarraysmentioning

confidence: 96%

Pathogen specific carbohydrate antigen microarrays: a chip for detection of Salmonella O-antigen specific antibodies

et al. 2007

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A Salmonella O-antigen microarray was developed by covalent coupling of oligosaccharide antigens specific for serogroups Salmonella enterica sv. Paratyphi (group A), Typhimurium (group B) and Enteritidis (group D). Antibodies were correctly detected in sera from patients with culture verified salmonellosis. High serogroup-specificity was seen with the disaccharide antigens. With the larger antigens, containing the backbone sequence Manalpha1-2Rhaalpha1-2Gal (MRG), common backbone-specific antibodies (O-antigen 12) were also detected. This is "proof of principle" that pathogen-specific carbohydrate antigen microarrays constitute a novel technology for rapid and specific serological diagnosis in either individual patients or larger sero-epidemiological and vaccine studies.

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New technology for regiospecific covalent coupling of polysaccharide antigens in ELISA for serological detection

Cited by 18 publications

References 14 publications

Development of a Bead Immunoassay To Measure Vi Polysaccharide-Specific Serum IgG after Vaccination with the Salmonella enterica Serovar Typhi Vi Polysaccharide

Development of a Bead Immunoassay To Measure Vi Polysaccharide-Specific Serum IgG after Vaccination with the Salmonella enterica Serovar Typhi Vi Polysaccharide

High molecular weight neoglycoconjugates for solid phase assays

Pathogen specific carbohydrate antigen microarrays: a chip for detection of Salmonella O-antigen specific antibodies

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