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Lupine ELISA Kit

INTENDED USE

Lupine ELISA kit is designed for analysing quantitative amounts of lupine (lupin and lupinus) in food (such as sausage, ketchup, potato products, juices and bakery products). This assay has a minimum analytical sensitivity limit of 0.2 ppm.

BACKGROUND

Lupine (also referred to as lupin or lupinus) belongs to the genus of legume family, which includes over 200 different species. The lupin seed contains a high fraction of proteins (approx. 35-45%) and these proteins (mainly α-, β- and γ-conglutin fractions) usually lead to allergic reactions because of their cross reactivity to allergens of peanut and soya. Lupine is frequently in dietary products, in particular for individuals that are allergic to milk, patients with celiac disease and vegetarians.

Due to high cross-reactivity, the following assay is suitable for measuring all commonly used lupine species. During many food production processes there are three sweet lupine species which are often used: Lupinus angustifolius, Lupinus luteus and Lupinus albus. The presence of lupine allergins in foods can lead to critical problems to people that are allergic to lupin. Even very low levels can result in an allergic reaction and in severe cases can lead to anaphylactic shock.

LUPINE ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Microtiter Plate: Coated with anti lupin antibodies.
  • Lupin Standards 1-5: Concentration 0, 2, 5, 15, 30 ppm.
  • Conjugate (Anti Lupin Peroxidase).
  • Substrate Solution (TMB).
  • Stop Solution (0.5 M H2SO4).
  • Extraction and sample dilution buffer (Tris)(10x).
  • Washing Solution (PBS + Tween 20)(10x Concentrate).
  • Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of lupin and lupinus derivatives current lupine ELISA kit was 0.2 ppm. The standard range for this assay is 2.0 – 30.0 ppm.

ASSAY CHARACTERISTICS

– Lupine (Lupin): ELISA
– Sensitivity: Limit of detection, LOD (0.02ppm), Limit of quantification, LOQ (2.0ppm)
– Intra-Assay Precision: 6 – 9%
– Inter-Assay Precision: 5 – 9%
– Linearity: 78 – 126%
– Cross Reactivity: Soy flour (unroasted)(0.07%), Soy lecithin (0.002%), Soy flour (roasted) (0.0009%), Lentil (0.0004%), Chickpea (0.0003%), Hazelnut (0.0002%). No cross reaction was detected for the following: Milk, Sunflower seeds, Cocoa, Egg, Pumpkin seeds, Potato, Wheat, Cashew, Leek, Rye, Peanut, Pea, Oats, Guar gum, Bean, Barley, Almond, Tomato, Rice, Pecan, Peach, Corn, Coconut, Cherry, Buckwheat, Brazil nut, Orange, Sesame, Walnut, Strawberry, Pork, Pistachio, Plum, Beef, Macadamia nut, Locust bean gum, Chicken, Chestnut.
– Recovery: Bread (113%), Croquette (111%), Orange juice (104%), Sausage (99%), Ketchup (98%).

REFERENCES

  1. Quantitative sandwich ELISA for the determination of lupine (Lupinus spp.) in foods. J Agric Food Chem. (2005) 53 (15): 5866-71. Holden L., et al.
  2. Characterization of IgE binding to lupin, peanut and almond with sera from lupin-allergic patients. Int Arch Allergy Immunol. (2008) 146 (4): 267-76. Holden L., et al.
  3. Validation and comparison of a sandwich ELISA, two competitive ELISAs and a real-time PCR method for the detection of lupine in food. Food Chem. (2013) 141 (1): 407-18. Ecker C., et al.
  4. Novel polyclonal-monoclonal-based ELISA utilized to examine lupine (Lupinus species) content in food products. J Agric Food Chem. (2007) 55 (7): 2536-42. Holden L., et al.
  5. Lupine-induced anaphylaxis. Ann Allergy Asthma Immunol. (1999) 83 (5): 406-8. Matheu V., et al.
  6. Sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of lupine residues in foods. J Food Sci. (2008) 73 (8): T135-40. Kaw C.H., et al.

ADDITIONAL INFORMATION

  • Full Name: Lupine ELISA Kit
  • Sample Type: Food (Bakery, Sausage, Potato, Ketchup, Juices)
  • Sensitivity: 0.2 ppm

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