title: Development and evaluation of a monoclonal antibody-based inhibition ELISA for the quantification of chymosin in solution.
abstract: Chymosin is the major enzyme of natural rennet, traditionally used in cheese making for its high milk-clotting activity. For technical reasons, an accurate characterization of rennet should include its total clotting activity and also its enzymatic composition. Monoclonal antibodies specific to chymosin were obtained from mice immunized with purified bovine chymosin, and an inhibition enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of chymosin in solution. No cross-reactivity was observed with other milk-clotting enzymes commonly used in cheese making. The limit of detection and limit of quantification were 125 and 400 ng/mL, respectively. The values of precision within and among runs were 7.23 and 7.39%, respectively, and satisfying recovery, from 92 to 119%, was found for spiked samples. The inhibition ELISA was successfully applied to commercial rennets, and the results were consistent with those obtained using the standard chromatographic method (IDF 110: A, 1987).
authors.affiliation: †INRA, UR342 Technologie et Analyses Laitières, F-39800 Poligny, France.
authors.forename: O
authors.initials: O
authors.lastname: Rolet-Répécaud
authors.affiliation: †INRA, UR342 Technologie et Analyses Laitières, F-39800 Poligny, France.
authors.forename: C
authors.initials: C
authors.lastname: Arnould
authors.forename: D
authors.initials: D
authors.lastname: Dupont
authors.affiliation: ∥ACTALIA, F-25620 Mamirolle, France.
authors.forename: S
authors.initials: S
authors.lastname: Gavoye
authors.affiliation: †INRA, UR342 Technologie et Analyses Laitières, F-39800 Poligny, France.
authors.forename: E
authors.initials: E
authors.lastname: Beuvier
authors.affiliation: †INRA, UR342 Technologie et Analyses Laitières, F-39800 Poligny, France.
authors.forename: C
authors.initials: C
authors.lastname: Achilleos
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D002417:Cattle; D002611:Cheese / Q000032:analysis*; D012085:Chymosin / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005503:Food Additives / Q000032:analysis*; D051379:Mice; D008892:Milk / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D005503:Food Additives; D012085:Chymosin
keywords: chymosin; inhibition ELISA; monoclonal antibody; rennet