Sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of lupine residues in foods

C. H. Kaw, S. L. Hefle, Stephen L Taylor

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Lupine has been increasingly used in food applications due to its high nutritional value and excellent functional properties. However, lupine provokes allergic reactions in susceptible individuals. The presence of undeclared lupine residues in foods can pose a serious health risk to lupine-allergic individuals. Therefore, the objective of this research was to develop a sandwich-type ELISA for the detection of lupine residues in foods. Lupine flour derived from Lupinus albus was used to immunize 3 rabbits and a sheep. Pooled lupine-specific antibodies were partially purified from the sera by ammonium sulfate precipitation. A sandwich lupine ELISA with a limit of quantification (LOQ) of 1 ppm was developed by utilizing the rabbit antisera as the capture reagent and the sheep antiserum as the detector reagent. The binding of the antigen-antibody complex was visualized by the addition of commercial rabbit antisheep IgG antibody labeled with alkaline phosphatase with subsequent addition of p-nitrophenyl phosphate substrate to produce a colored product for quantification. Minor cross-reactivity was observed with soy (Glycine max) and black bean (Castanospermum australe). The performance of the lupine ELISA was evaluated in reference food standards (beef frankfurter and apple cinnamon muffin) and laboratory-prepared cooked frankfurters and corn muffins. The mean percent recovery for lupine spiked-frankfurters and corn muffins were 108.4% ± 8.8% and 103.1% ± 11.5%, respectively. The sandwich-type lupine ELISA developed in this study provides food manufacturers and regulatory agencies with an effective analytical tool to detect and quantify lupine residues in processed foods.

Original languageEnglish (US)
JournalJournal of Food Science
Volume73
Issue number8
DOIs
StatePublished - Oct 1 2008

Fingerprint

Lupinus
sandwiches
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
Food
muffins
hot dogs
rabbits
Rabbits
Castanospermum
antiserum
Castanospermum australe
Zea mays
Immune Sera
Sheep
black beans
Cinnamomum zeylanicum
sheep
antigen-antibody complex
antibodies

Keywords

  • Allergen control
  • ELISA
  • Immunoassay
  • Lupine
  • Lupine allergy

ASJC Scopus subject areas

  • Food Science

Cite this

Sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of lupine residues in foods. / Kaw, C. H.; Hefle, S. L.; Taylor, Stephen L.

In: Journal of Food Science, Vol. 73, No. 8, 01.10.2008.

Research output: Contribution to journalArticle

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