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GrainGenes Reference Report: JCS-30-267

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Reference	JCS-30-267
Title	Antibodies to N-terminal peptides of low M(r) subunits of wheat glutenin. I. Characterisation of the antibody response
Journal	Journal of Cereal Science
Year	1999
Volume	30
Pages	267-281
Author	Sissons MJ Blundell MJ Hill AS Skerritt JH
Abstract	The relationship between allelic composition of the low molecular mass glutenin subunits (LM(r)GS) and dough properties is poorly understood. Differentiating the LM(r)GS on the basis of their N-terminal sequence type may provide an important alternative in understanding the relationship. Polyclonal and/or monoclonal antibodies were produced using synthetic peptides corresponding to each of the seven N-terminal amino acid sequence types of the B- and C- LM(r)GS, namely SHIPGLERPS-, METSHIPGL-, METSRVPGL-, METSCIPGL-, METRCIPGL-, NMQVDPSGQVQ-(gamma-type) and VRVPVPQLQP-(alpha-type). Each of the polyclonal antisera recognised both the corresponding peptide and LM(r)GS. For monoclonal antibodies, the proportion of hybridoma clones that produced antibody which recognised either the peptide or LM(r)GS varied between 1 and 88%. However, antibodies from only 4% of antibody-secreting stable cell lines recognised both the peptide immunogen and intact LM(r)GS. Using ELISA, the majority of the antibodies cross-reacted with related synthetic peptides corresponding to more than one N-terminal LM(r)GS sequence, although several of these bound small groups of LM(r)GS on immunoblots. Different polyclonal antisera prepared to a given immunogen exhibited similar patterns of subunit recognition on immunoblots. Monoclonal antibodies prepared to the same immunogen exhibited a variety of patterns, although each of the antibodies specific for a particular peptide or combination of peptides on ELISA recognised a similar pattern of LM(r)GS on immunoblots. For each sequence type, polyclonal or monoclonal antibodies specific for individual N-terminal sequences were identified. These probes may be useful tools to determine whether the type and amount of each N-terminal sequence is correlated with dough properties
Keyword	[ Hide all but 1 of 28 ] amino acid amino acid sequence antibody antibody formation antiserum cell lines clone cross reaction dough dough properties elisa epitopes glutenin glutenin subunit immune serum immunoblotting low m(r) subunits m(r) monoclonal antibodies n-terminal amino acid sequence n-terminal peptides peptide probes recognition sequence synthetic peptides wheat flour wheat glutenin

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