Lysozyme ELISA Kit

Full Name: Lysozyme ELISA Kit
Sample Type: Food (Wine, Cheese)
Sensitivity: 2.0 ppb

BACKGROUND

Lysozyme (1,4-β-N-acetylmuramidase) is an antimicrobial enzyme that prevents bacterial infections. It is produced by animals as part of the innate immune system. Reduced levels have also been linked to bronchopulmonary dysplasia in newborns. It is abundant in secretions such as saliva, mucus, tears and human milk. In humans, erythrocyte lysosomal hydrolase activity is about 50% of control (estimated at 110,000 units/h/ml) in the circulation. The enzyme has been proposed as a possible biomarker for lung function or lung disease.

In humans, hen’s egg (also called Gallus gallus) provides a rich food source, where the proteins of the egg yolk display minor allergenicity and the egg white proteins are known to be allergenic. For example, Isoallergenic hen egg protein (IHEP) is composed of α- and β-subunits with a molecular weight of 58.3 kDa. In animals, the presence of chicken proteins is not usually a problem unless they are allergic to poultry vaccines which might be around 1%. Chickens that have been vaccinated with an egg protein immunological adjuvant show no clinical signs for at least forty-eight hours post vaccination due to the medication’s effect on the chicken’s immune system.

An individual that is allergic, even small amounts of consumption of lysozyme can cause an allergic reactions and in some case could lead to severe anaphylactic shock. In addition to ovotransferrin, livetin, ovalbumin and ovomucoid, lysozyme represents a very important allergen. Allergenicity is often related to the cell wall of the allergens. Lysozyme is a glycoprotein and it’s allergenic properties depend on its interaction with sialic acid or sulfate. It is used as a topical agent to treat keratoconjuctivitis sicca, also known as dry eye syndrome. It is found in human tears which come in contact with the ocular surface to fight against microbial infection and reduce inflammation.

INTENDED USE

Lysozyme ELISA kit is intended for detecting quantitative concentrations of lysozyme residues in food (such as wine or cheese products). This assay has a minimum analytical sensitivity limit of 2.0 ppb.

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-lysozyme antibodies.
Lysozyme standards 1-5: Concentration 0, 25, 50, 100, 250 ppb.
Conjugate (Anti-Lysozyme-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 lysozyme residues using current lysozyme ELISA kit was 2.0 ppb. The standard range for this assay is 25.0 – 250.0 ppb.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (2.0ppb), Limit of quantification, LOQ (25.0ppb)
– Cross Reactivity: Egg white protein, total (2.2%), Ovalbumin, Conalbumin, Ovomucoid (< 0.0001%), Chicken (< 0.000006%), Cashew (< 0.000005%). No cross reaction was detected for the following: Adzuki, Cow’s milk, Pecan, Almond, Cumin, Pepper, Apricot, Duck, Pine seed, Barley, Fenugreek, Pistachio, Bean (white), Gliadin, Poppy seed, Beef, Goat’s milk, Pork, Bovine gelatine, Guar gum, Potato, Brazil nut, Hazelnut, Prawn (cooked), Buckwheat, Kidney bean, Prawn (raw), Caraway, Kiwi, Pumpkin seed, Carob gum, Lamb, Rice, Carrot, Lentil, Rye, Cayenne, Linseed, Saccharose, Celery, Lupin, Sesame, Cherry, Macadamia, Shrimps, Chestnut, Mustard, Soy, Chia, Nutmeg, Soy lecithin, Chickpea, Oats, Split peas, Chili, Onion, Sunflower seeds, Cocoa, Paprika, Tomato, Coconut, Pea, Turkey, Cod, Peach, Walnut, Corn, Peanut, Wheat.
– Intra-Assay Precision: 2 – 6%
– Inter-Assay Precision: 2 – 3%
– Linearity: 87 – 98%
– Recovery: Wine (99%), Cheese (85%).

REFERENCES

Evaluation of the efficiency of enological procedures on lysozyme-depletion in wine by an indirect ELISA method. J Agric Food Chem. (2014) 62 (26): 6247-53. Carstens C., et al.
Sequential separation of lysozyme, ovomucin, ovotransferrin, and ovalbumin from egg white. Poult Sci. (2014) 93 (4): 1001-9. Abeyrathne E.D., et al.
Development of an ELISA for quantifying lysozyme in hen egg white. J Agric Food Chem. (2005) 53 (7): 2379-85. Vidal M.L., et al.
Detection of hen’s egg white_lysozyme in food: comparison between a sensitive hplc and a commercial ELISA method. Commun Agric Appl Biol Sci. (2007) 72 (1): 215-8. Kerkaert B. and De Meulenaer B.
The antimicrobial peptide-lysozyme is induced after multiple trauma. Mediators Inflamm. (2014) 2014: 303106. Klüter T., et al.

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