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A sensitive monoclonal‐antibody‐based test for gluten detection: Choice of primary and secondary antibodies
Author(s) -
Skerritt John H.,
Diment John A.,
Wrigley Colin W.
Publication year - 1985
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.2740361015
Subject(s) - monoclonal antibody , chemistry , antibody , horseradish peroxidase , primary and secondary antibodies , coeliac disease , peroxidase , wheat gluten , gluten , biochemistry , prolamin , tissue transglutaminase , chromatography , enzyme , biology , storage protein , immunology , medicine , disease , pathology , gene
Of a series of monoclonal antibodies prepared to cereal proteins, two antibodies with specificity for low‐mobility, heat‐stable prolamins in wheat and related cereals were investigated as possible probes for a test for gluten in cooked or processed foods. Urea‐based solvents were found to be superior to isopropanol or sodium dodecylsulphate extractants in allowing sensitive detection of trace amounts of prolamins. The antibodies detected bread and durum wheat and rye prolamins most strongly, followed by barley then oats; detection of maize and rice was quite weak. This selectivity is suitable for a test for prolamins toxic to coeliac‐disease patients. Several enzyme‐labelled second antibodies, for detection of monoclonal antibody bound to cereal protein, were found to be unsuitable reagents since an appreciable fraction of the second antibodies bound directly to the cereal proteins. Sensitive, artifact‐free detection of antibody binding could be performed using the peroxidase‐antiperoxidase technique or by direct conjugation of horseradish peroxidase to the monoclonal antibodies.