Development of an Indirect Competitive ELISA for Flumequine Residues in Raw Milk Using Chicken Egg Yolk Antibodies

• Published in: Journal of agricultural and food chemistry Vol. 52; no. 16; pp. 4975 - 4978
• Main Authors: Van Coillie, Els, De Block, Jan, Reybroeck, Wim
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 11.08.2004
• Subjects: Animals; antibodies; antimicrobial agents; Biological and medical sciences; bovine serum albumin; Chickens - immunology; drug residues; egg yolk; Egg Yolk - immunology; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Fluoroquinolones; food contamination; Food industries; Fundamental and applied biological sciences. Psychology; Immunoglobulins; Meat and meat product industries; Milk - chemistry; milk analysis; Milk and cheese industries. Ice creams; ovalbumin; Quinolizines - analysis; Quinolizines - immunology; quinolones; raw milk; Sensitivity and Specificity; Poultry; Meat product; Raw milk; Meat animals; Egg product; Development; Flumequine; IgY; Aves; Egg yolk; Immunoglobulins; Farming animal; Antibody; Dairy product; milk; Enzyme immunoassay; Chicken meat; Vertebrata; Residue; chicken immunoglobulins; Chicken; fluoroquinolones; Detection; ELISA assay; ELISA
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf049593d

To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA−flumequine) and to cationized ovalbumin (cOVA−flumequine). For the immunization of chickens, cBSA−flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA−flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 μg/kg). No cross-reactivity was noticed with various related quinolones. Keywords: Flumequine; fluoroquinolones; milk; IgY; chicken immunoglobulins; ELISA