Universidad De Sevilla

Biomedal, S.l.

Angel Cebolla Ramirez

Carolina Sousa Martin

Isabel Comino Montilla

Ana Real Calderon

Santiago Vivas Alegre

Jesús M Sanz Morales

Beatriz Maestro Garcia-Donas

Miguel Arevalo Rodriguez

Isabel Velasco Umpierrez

Moisés Alvarez Maqueda

Jesús Miguel Sanz Morales

The present invention fitted in the medical-clinical sector, shows a method for monitoring the ingestion of gluten by measuring protein/gluten peptides present in fecal samples with antibodies against immunogenic peptides resistant to gastrointestinal digestion. The presence or absence of these immunogenic peptides is controlled by immunological assays based on antibodies reactive against immunogenic gluten peptides that are resistant to proteolysis. These assays may be quantitative techniques ELISAs, or qualitative as rapid immunochromatographic assays, immunoblots, etc. These measures may also be applied to verify compliance with the gluten-free diet, to improve diagnosis in cases of refractory or severe symptoms of celiac disease in which a gluten-free diet is supposedly being respected, or to clinical research on the effectiveness of enzymatic therapies related with prolamin detoxification.

Determination of levels of immunogenic gluten peptides in human samples

The present invention, fitted in the medical-clinical sector, shows a method for monitoring the ingestion of gluten by measuring protein/gluten peptides present in fecal samples with antibodies against immunogenic peptides resistant to gastrointestinal digestion. The presence or absence of said immunogenic peptides is controlled by immunological assays based on reactive antibodies against immunogenic gluten peptides that are resistant to proteolysis. These assays may be quantitative techniques as ELISAs, or qualitative as rapid immunochromatographic assays, immunoblots, etc. These measures may also be applied to verify compliance with the gluten-free diet, to improve diagnosis in cases of refractory or severe symptoms of celiac disease, in cases in which a gluten-free diet is supposedly being respected, or to clinical research on the effectiveness of enzymatic therapies related with prolamin detoxification.

The object of the present invention is a procedure for the distribution, separation and purification in aqueous solution of recombinant proteins, based on the utilization of polypeptides with choline affinity. The invention is based on a phenomenon consisting of that two aqueous solutions with determinated components can be mixed, being distributed finally in two phases with different density. The fusionated proteins to said polypeptides with choline affinity are preferably located in one of the phases, while most of the cell extract proteins tend to go to the opposite phase. After a series of washings for removing the rest of the not desired material, this location can be inverted through the addition of a soluble molecule with affinity by the polypeptide fusionated to the protein of interest. This procedure allows modulating at convenience the presence of the protein or polypeptide of interest in one phase or another, possibiliting its purification with a high yield and purity grade. The invention represents an economic and scalable way of recombinant protein separation labelled preferably with choline-binding domains.

Methods for separating recombinant proteins in aqueous two-phase systems

The invention relates to a method for preserving peptides or proteins over long periods of time. The invention also relates to the use of the preservation method for the transport of labeled peptides or proteins without requiring refrigeration, as well as to a set of tools for implementing said method.

Peptide and protein preservation method

Growing community of inventors

Sevilla, Spain

Angel Cebolla Ramirez