Cereal Chem. 73 (5):650-657  |  VIEW ARTICLE

Analytical Techniques and Instrumentation

Wheat Dough Extensibility Screening Using a Two-Site Enzyme-Linked Immunosorbent Assay (ELISA) with Antibodies to Low Molecular Weight Glutenin Subunits (1).

J. L. Andrews (2) and J. H. Skerritt (2,3). (1) Presented in part at the AACC 79th Annual Meeting, Nashville, TN, October 1994. (2) CSIRO Division of Plant Industry, North Ryde, NSW 2113 and Canberra, ACT 2601, Australia. (3) Corresponding author. CSIRO Division of Plant Industry, GPO Box 1600, Canberra ACT 2601 Australia. Fax + 61 6 246 5351/ 5000; Phone + 61 6 246 5350; E-mail: <kerritt@pi.csiro.au> Accepted June 3, 1996. Copyright 1996 by the American Association of Cereal Chemists, Inc. 

The relationships between total glutenin subunit content, low molecular weight glutenin subunit (LMW-GS) content, and dough extensibility were examined with several sets of flours, in which the overall correlations between flour protein content and extensibility were poor. The potential for monoclonal antibodies with specificities for various LMW-GS groups to screen for differences in dough extensibility was assessed using a two-site (sandwich) enzyme-linked immunosorbent assay (ELISA), following extraction of LMW-GS using 60% ethanol and 10mM dithiothreitol. A range of solid-phase bound (immobilized) antibodies with specificities for different LMW-GS were combined with two labeled (detection) antibodies: 21817, which bound a cluster of B-LMW-GS, and 23724, which bound both low molecular weight glutenin subunits (HMW-GS) and B-LMW-GS. Significant correlations were obtained with each flour set using the more specific detection antibody, 21817, although the specificity of the immobilized antibody was less important. Binding of the antibody combinations that related to extensibility correlated with LMW-GS content measured using reversed-phase high-performance liquid chromatography (RP-HPLC). The antibody binding, RP-HPLC, and extensibility correlations were of the order r = 0.4-0.7, which suggested that variation in total LMW-GS content accounted for only part of the variation in extensibility between flours. It is possible that interactions between several factors, including allelic composition of HMW-GS and LMW-GS as well as total LMW-GS content may determine dough extensibility.