Enzyme-linked immunosorbent assay for ileal human goblet cell mucin

Roberten, Anthony M.; Dalzell, Howard; Harding, Carol A.; Pearce, Steven; Ram, Satyendra P.; McKay, E.J.; Wesley, Alison

doi:10.1016/0009-8981(88)90321-x

Cited by 3 publications

(2 citation statements)

References 17 publications

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“…This was also described in the literature for bovine [21], human [36] and rabbit [37] gastro-intestinal glycoprotein. This suggests the presence of common antigenic determinants among these glycoproteins both within and between animal species.…”

Section: Hyperimmune Plasma Characterization and Elisa Assay Set Upmentioning

confidence: 57%

“…This range was intermediate between other direct ELISA assays for human (2 to 15 ng·mL -1 ) [36], rat (0 to 20 ng·mL -1 ) [35] or preruminant calf (20 to 640 ng·mL -1 ) [21] small intestinal glycoprotein. The ELISA intraand inter-plate variabilities were very similar between porcine (present work) and bovine glycoprotein (previous work, [21] ).…”

Section: Hyperimmune Plasma Characterization and Elisa Assay Set Upmentioning

confidence: 75%

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Estimation of ileal output of gastro-intestinal glycoprotein in weaned piglets using three different methods

et al. 2004

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-Mucin is the main constituent of gastrointestinal mucus and is responsible for its physicochemical and physiological properties. Previous studies have suggested that this glycoprotein represents a major component of undigested endogenous protein at the ileum. The aim of the study was to estimate the ileal output of this glycoprotein using three methods: direct ELISA, hexosaminebased method and ethanol precipitation. For setting up the ELISA assay, the glycoprotein was isolated from intestinal mucus scraping by cesium chloride density gradient ultracentrifugation and a rabbit hyperimmune plasma was raised against the purified glycoprotein. Ileal outputs of hexosamine and glycoprotein were measured in weaned piglets fed a control diet (C) based on casein or diets which contained 50% crude protein supplied by white (WCP) or black (BCP) chickpea. The hexosamine output was higher (P < 0.05) with the WCP diet (2.3 and 1.5 g·kg -1 of dry matter intake for glucosamine and galactosamine, respectively) than with diet C (1.1 and 0.7 g·kg -1 of DMI). The hexosamine-based and ethanol precipitation methods, but not the ELISA, showed significant differences between the diet treatments (P < 0.05). Although hexosamine-based and ethanol precipitation methods for the estimation of ileal glycoprotein appeared to be more satisfactory than the developed ELISA to display diet effects in this study, it remains to be determined whether the higher glycoprotein data variability observed with ELISA reflects the actual biological variability of the phenomenon or not.small intestine / methods / glycoprotein / piglet

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Section: Hyperimmune Plasma Characterization and Elisa Assay Set Upmentioning

confidence: 57%

Section: Hyperimmune Plasma Characterization and Elisa Assay Set Upmentioning

confidence: 75%

Estimation of ileal output of gastro-intestinal glycoprotein in weaned piglets using three different methods

et al. 2004

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In vitro adsorption of a hydrophobic mutagen to gastrointestinal mucus glycoprotein (mucin) and dietary fibre

Ferguson

Harris

et al. 1992

Chemico-Biological Interactions

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Calf Intestinal Mucin: Isolation, Partial Characterization, and Measurement in Ileal Digesta with an Enzyme-Linked Immunosorbent Assay

Montagné¹,

Toullec²,

Lallès³

2000

Journal of Dairy Science

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The aim of this study was to develop a specific ELISA for calf intestinal mucin to quantify its contribution to ileal endogenous losses. Mucin was isolated from intestinal scrapings by cesium chloride density gradient ultracentrifugation. The isolated mucin had a high concentration of glutamic and aspartic acids, threonine, and serine (13.2, 11.2, 9.6, and 9.2 mol % of total amino acids assayed, respectively). The carbohydrates present were (mol % of total hexoses): galactose 42.1, N-acetylglucosamine 24.1, N-acetylgalactosamine 23.6, fucose 4.7, mannose 3.1, and sialic acids 2.4. Amino acids and carbohydrates represented 52.6 and 47.4% of the mucin by weight, respectively. A rabbit hyperimmune plasma was raised against purified mucin and used to set up an ELISA. The linear range of this assay was 20 to 640 ng/ml. The plasma cross-reacted with calf abomasal and colonic mucin. It showed no cross-reactivity with nonmucin components and no reactivity with intestinal mucin from other animal species except for the rat. Mucin accounted for approximately 3.5% of the dry matter output at the ileum of calves fed a substitute milk based on skim milk powder. This represented a flow of 3.4 g of mucin/kg of dry matter intake. Mucin flow increased when dietary protein was provided by cow's colostrum. Finally, the developed assay is a suitable tool to investigate the impact of dietary factors on the flow of mucin along the gut of preruminant calves.

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Enzyme-linked immunosorbent assay for ileal human goblet cell mucin

Cited by 3 publications

References 17 publications

Estimation of ileal output of gastro-intestinal glycoprotein in weaned piglets using three different methods

Estimation of ileal output of gastro-intestinal glycoprotein in weaned piglets using three different methods

In vitro adsorption of a hydrophobic mutagen to gastrointestinal mucus glycoprotein (mucin) and dietary fibre

Calf Intestinal Mucin: Isolation, Partial Characterization, and Measurement in Ileal Digesta with an Enzyme-Linked Immunosorbent Assay

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