Rmf Dictagene S.a.

Anergis S.a.

Christophe Dominique Reymond

Nicolas Joseph Fasel

Francois Spertini

Giampietro Corradin

Mario Roggero

Alexander Kettner

Contiguous overlapping peptides (COPs) for the treatment of allergic patients by Specific Immunotherapy (SIT) are provided from the sequence of the major allergen of ragweed pollen Amb a 1. Such peptides while providing all potential T cell epitopes are devoid of the three-dimensional structure of the original allergen, therefore reducing their ability to bind IgE.

Contiguous overlapping peptides for treatment of ragweed pollen allergy

Contiguous overlapping peptides (COPs) for the treatment of allergic patients by Specific Immunotherapy (SIT) are provided from the sequence of the major allergen of birch pollen Bet v 1. Such peptides while providing all potential T cell epitopes are devoid of the 3D structure of the original allergen, therefore reducing their ability to bind IgE.

Contiguous overlapping peptides for treatment of birch pollen allergy

The invention relates to the use of the regulatory subunit (R) of the cAMP dependent protein kinase (PKA) from for cAMP detection. It includes constructs for expression of the R-subunit in and fusion to green fluorescent proteins (GFP). Fluorescence energy transfer is used as a way to monitor cAMP binding, either by using fluorescently labelled cAMP or cGMP, or by using mutant GFPs with modified absorption and emission spectra. FRET changes upon cAMP binding will allow measurement of cAMP level either in vitro or within living cells.

Use of the regulatory subunit of the camp dependent protein kinase (PKA) from dictyostelium for camp measurements

High expression levels and simple downstream processing are essential for the production of recombinant proteins at low cost. We report here a new expression vector which allows production of fusion proteins in . We made use of the ability of Discoidin I to bind Sepharose-4B to set up a nearly single step purification procedure. The Discoidin I coding region was fused to several forms of the malaria parasite CSP gene in a Dictyostelium expression vector, allowing intracellular accumulation as well as partial secretion via a pathway unrelated to the endoplasmic reticulum and Golgi. The fusion proteins present in cell extracts were affinity-purified over Sepharose-4B columns. Addition of a signal peptide allowed endoplasmic reticulum targeting and glycosylation of the fusion protein. Inclusion of a thrombin cleavage site allowed to cleave Discoidin from the CSP protein. The use of stable and low cost Sepharose 4B as affinity matrix should allow large-scale preparations.

Expression vector for use in a one-step purification protocol

A recombinant polypeptide comprising a polypeptide which has added thereto a glycoxyl-phosphatidylinositol anchor structure, said polypeptide demonstrating a greater immune response than the corresponding polypeptide without the anchor. The polypeptide may be a parasite antigen such as a Plasmodium falciparum polypeptide. Recombinant vectors comprising the DNA sequence encoding the polypeptide with a glycolipid anchor addition sequence and host cells based thereon are also disclosed as well as vaccines and methods of inducing an immune response based on the polypeptides.

Modified polypeptides for enhanced immunogenicity

Preparation of purified (poly)peptides

The invention relates to recombinant DNA molecules comprising a Dictyostelium discoideum homologous promoter region, a heterologous DNA sequence with a Dictyostelium discoideum homologous peptide sequence capable of functioning as a leader peptide sequence positioned upstream thereof and in proper reading frame therewith, and a Dictyostelium discoideum homologous termination region, the heterologous DNA sequence encoding the desired functional polypeptide or intermediate thereof, said DNA sequence being positioned downstream from the promoter region and said termination region being positioned downstream from the DNA sequence. The invention also provides recombinant dictyostelid hosts as well as methods for preparing recombinant DNA molecules of the invention and for expressing recombinant polypeptides encoded by said recombinant DNA molecules in dictyostelid hosts. The invention in particular relates to the expression of the CS protein of Plasmodium falciparum.

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Prilly, Switzerland

Christophe Dominique Reymond