Production of a horse‐specific monoclonal antibody and detection of horse meat in raw meat mixtures by an indirect ELISA

Abstract

A stable hybridoma cell line (DD3) has been produced secreting a monoclonal antibody specific for horse muscle proteins. The DD3 monoclonal antibody (mAb) did not show significant cross-reactivity when tested against beef, chicken, pig and soya proteins or bovine caseins, gelatine and bovine serum albumin. The DD3 mAb was used in an indirect ELISA format for the detection of defined amounts of horse meat (10-500 g kg-') in beef meat mixtures. Immunorecognition of monoclonal antibodies adsorbed to horse meat adsorbed onto the ELISA plate was made with rabbit anti-mouse immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymic conversion of the substrate gave clear optical density differences when assaying mixtures of minced beef containing different amounts of horse meat.

Monoclonal Antibodies to Porcine Thermal‐Stable Muscle Protein for Detection of Pork in Raw and Cooked Meats

“…Hybridoma techniques, developed by Köhler and Milstein (1975), enabled continuous production of MAbs with defined specificity. MAbs have been applied to ELISAs for raw meat identification (Martin et al, 1991;Garcia et al, 1994;Morales et al, 1994;Billett et al, 1996). However, MAbs have not been developed for detection of species adulteration in cooked meat products.…”

Detection of Horse Meat Contamination in Raw and Heat-Processed Meat Products

“…The 96-well microplate format test kit marketed by ELISA Technologies Inc. (Gainesville FL, USA), suffers from similar limitations as it is also based on pAbs. Although Garcia and others 17 noncompetitive indirect ELISA (iELISA) for the detection of horse meat, this approach is again applicable only to raw meats. The objective of this study was, therefore, to (1) develop horse selective monoclonal antibodies (mAbs) using thermally stable horse muscle proteins as the immunogen and, consequently, (2) develop a suitable mAb-based ELISA for the quantitative detection of horse meat in both raw and heatprocessed ground meat samples.…”

“…The 96-well microplate format test kit marketed by ELISA Technologies Inc. (Gainesville FL, USA), suffers from similar limitations as it is also based on pAbs. Although Garcia and others have reported the use of mAbs as probes in noncompetitive indirect ELISA (iELISA) for the detection of horse meat, this approach is again applicable only to raw meats.…”

Identification of the Clam Species Ruditapes decussatus (Grooved Carpet Shell), Venerupis pullastra (Pullet Carpet Shell), and Ruditapes philippinarum (Japanese Carpet Shell) by PCR-RFLP

“…In contrast to the numerous analytical methods that have been developed for meat (Garcı ´a et al, 1994;Matsunaga et al, 1999), milk (Anguita et al, 1995;Dennis, 1996), or fish (Carrera et al, 1999a(Carrera et al, ,1999bRehbein et al, 1999) species identification, molluscs have received little attention so far.…”