Cereal Chem 68:467-474 | VIEW
A Simple Antibody-Based Test for Dough Strength. I. Development of Method and Choice of Antibodies.
J. H. Skerritt. Copyright 1991 by the American Association of Cereal Chemists, Inc.
A monoclonal antibody-based enzyme-immunoassay method was developed to predict dough strength and mixing properties from the analysis of small samples of wheat flour or whole meal. The choice of antibody, grain protein extractant, and extraction technique were important. Solvents such as alkali or acid extracted. Quality-related differences in antibody binding to grain proteins extracted using sodium dodecyl sulfate (SDS) with a reducing agent (dithiothreitol [DTT]) were noted. Proteins could be extracted by vortex mixing; ultrasonic or high-frequency homogenization were inferior. Quality-related differences could be measured using grain extracted a single time, although the SDS-DTT extractant provided greater differentiation when a simple preextraction step was performed using SDS. The specificities of the antibody combinations were assessed using gluten protein fractions and genetic wheat stocks, including Chinese Spring aneuploids and near-isogenic wheats that lack certain high and low molecular weight glutenin subunits and gliadins. The best quality differentiation was obtained using antibody combinations that bound to chromosome 1D-encoded high molecular weight glutenin subunits. However, these antibodies bound to glutenin subunits both from varieties having allelic subunits 1Dx5 and 1Dy10 and from varieties having allelic subunits 1Dx2 and 1Dy12, although greater binding to proteins was seen from extracts of wheat flours that yielded stronger doughs.