Gonzalo Anguita

Immunostick ELISA for Detection of Cow's Milk in Ewe's Milk and Cheese Using a Monoclonal Antibody against Bovine β-Casein

An immunostick enzyme-linked immunosorbent assay (ELISA) has been developed for the rapid detection of cows milk in ewes milk or cheese. The assay uses a monoclonal antibody (AH4) produced against bovine β-casein for the detection of cows milk or cheese bound to the paddles of immunostick tubes. This immunostick ELISA allows the visual identification of ewes milk containing more than 1% of cows milk or cheese samples containing more than 0.5% of cows cheese.

A competitive enzyme-linked immunosorbent assay for detection of bovine milk in ovine and caprine milk and cheese using a monoclonal antibody against bovine beta-casein.

A competitive ELISA (enzyme-linked immunosorbent assay) was performed to detect and quantify bovine milk in ovine and caprine milk and cheese using a monoclonal antibody (AH4 MAb) against bovine beta-casein. Ovine or caprine milk and cheese containing bovine milk were added simultaneously with the AH4 MAb to the wells of a microtiter plate that had been previously sensitized with commercial bovine beta-casein. The bovine caseins in milk or cheese samples compete with the bovine beta-casein bound to the plate for the AH4 MAb binding sites. Further immunorecognition of AH4 MAb bound to the bovine beta-casein immobilized onto the plate was attained with rabbit anti-mouse immunoglobulin conjugated to peroxidase. Subsequent enzymic conversion of the substrate showed clear differences in absorbance values during assay of mixtures of ovine and caprine milk and cheese containing various amounts of bovine milk. The competitive ELISA developed in this work allows the quantitative detection of bovine milk in ovine and caprine milk and cheese samples in the range of 0.5 to 25% of substitution.

Detection and quantification of goat's cheese in ewe's cheese using a monoclonal antibody and two ELISA formats

A stable hybridoma cell line (B2B) has been produced secreting a monoclonal antibody (MAb) specific for the s αs2-casein of goats. The MAb B2B was used in two enzyme-linked immunosorbent assays (ELISA) formats for the detection and quantification of the presence of goats cheese in ewes cheese samples. In the indirect ELISA format the limit of detection was 1–25% (w/w) substitution of ewes cheese samples by goats cheese. Afterwards, a competitive indirect ELISA was successfully developed for the detection of 0.5 to 25% (w/w) of goats cheese in ewes cheese samples. This competitive indirect ELISA is a very sensitive assay, can be performed in less than 5 h and is not influenced by the ripening process in cheese.

Use of a monoclonal antibody and two enzyme-linked immunosorbent assay formats for detection and quantification of the substitution of caprine milk for ovine milk

A stable hybridoma cell line (B2B) has been produced that secretes a monoclonal antibody (MAb) specific for goats milk αS2-casein. The MAb B2B was used in two enzyme-linked immunosorbent assay (ELISA) formats for the detection and quantification of the presence of goats milk in ewes milk. In the indirect ELISA format the limit of detection was 0.5 to 15% (vol/vol) substitution of goats milk for ewes milk. Afterwards, a competitive indirect ELISA was successfully developed for the detection of 0.25 to 15% (vol/vol) of goats milk in ewes milk. This competitive indirect ELISA is a very sensitive assay; it can be performed in less than 5 h and is not influenced by the heat treatment of milk.

Antibody‐based analytical methods for meat species determination and detecting adulteration of milk

The food industry must provide consumers with products which are innocuous, genuine, desirable and nutritive. To assure consumers that food has these characteristics, it is convenient to have appropriate controls of their hygiene state and to detect fraudulent practices, both of which are possible using adequate analytical techniques. However, the analysis of food has its own limitations such as specificity, reproducibility, sensitivity, economy, ease of use and standardization. Immunological techniques have objective advantages which merit their development and further use in the food industry. It has been widely demonstrated that immunological methods can be used in the food industry as reliable analytical techniques. Antibody‐based analytical methods for determining meat species and detecting adulteration of milk have been developed. We review the efforts made to develop immunological reactions and techniques to detect fraudulent replacements of foods. The selection of antigens as immunogens, the devel...