The use of anti-soya globulin antisera in investigating soya digestionin vivo
✍ Scribed by Perez, M Dolores; Mills, E?N Clare; Lambert, Nigel; Johnson, Ian T; Morgan, Michael R?A
- Book ID
- 101226294
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 398 KB
- Volume
- 80
- Category
- Article
- ISSN
- 0022-5142
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✦ Synopsis
Polyclonal antisera have been developed which recognise the soya globulins glycinin and b-conglycinin. Their binding to proteolysed and processed globulins has been characterised with a view to using the antisera as probes to investigate the fate of soya globulins during digestion. Trypsinolysis reduced the immunoreactivity of b-conglycinin but increased that of glycinin by threefold, as determined by enzyme-linked immunosorbent assay (ELISA). However, both antisera recognised trypsinolysis products poorly by immunoblotting. These data are consistent with the fact that the anti-glycinin antiserum was raised to digested glycinin and would therefore recognise both intact and proteolytic intermediates of glycinin. The latter would be disrupted into component polypeptides by sodium dodecyl sulphate±polyacrylamide gel electrophoresis and hence would only be present in the ELISA and not after blotting. The antibody preparations were used to investigate the fate of soya globulins during in vivo digestion of a processed soya ingredient in rats. Both glycinin and b-conglycinin were digested rapidly, intact globulins disappearing from the gastrointestinal tract 3 h after dosing. Immunoreactive globulins were found in gut contents and associated with gut tissues but appeared to be in a semi-intact form, probably comprising proteolytic intermediates. This study demonstrates the usefulness of antibody methods in following the digestion of individual components in a complex mixture, such as is found in food. The availability of antibody preparations which recognise heat-stable epitopes will open the way for investigations into the effects of pre-treatments, such as cooking, which are pertinent to human consumption of legumes.