Roche Diagnostics Operations, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-La Roche Ag

Peter Schaarschmidt

Christian Scholz

Elke Faatz

Urban Schmitt

Alfred Engel

Barbara Upmeier

Herbert Andres

Toralf Zarnt

Ralf Bollhagen

Ariuna Bazarsuren

Hartmut Duefel

Alexander Riedel

Werner Stock

Christian Schotz

The invention concerns a fusion polypeptide including several molecules of folding helper polypeptides, including one multimerization domain, in particular Skp, and at least one molecule of SlyD or SlpA, wherein no further target polypeptide sequences are fused to the fusion polypeptide. The invention further concerns an immunoassay and the use of the fusion polypeptide in an immunoassay for reduction of interferences or minimizing false positive results or for stabilizing proteinaceous assay reagents. Further the invention concerns a reagent kit for use in an immunoassay comprising the fusion polypeptide.

Chaperone-chaperone fusion polypeptides for reduction of interference and stabilization of immunoassays

The invention relates to a method for detecting antibodies against the TpN17 antigen ofin an isolated sample wherein a peptide sequence oflipoprotein 15 (VcLp15) or a partial sequence thereof is used as a reagent for reduction of interference, i.e. for minimizing false positive results. In addition the invention relates to fusion polypeptides comprising a VcLp15 peptide sequence and a chaperone, to their use as an additive in an immunoassay for said reduction of interferences and for minimizing false positive results and to a reagent kit for detecting antibodies againstantigens in an isolated sample comprising a TpN17 antigen and said VcLp15-chaperone fusion polypeptide.

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The invention concerns soluble variants ofantigen 47 (TpN47 antigen) comprising at least domain B, or at least domains A and B, optionally domain D of the complete TpN47 protein molecule with the proviso that all antigens lack domain C (amino acid residues 224 to 351) of TpN47. The Tpn47 antigens can be fused to a chaperone. Moreover, the invention covers DNA encoding the antigens, a method of producing these antigens as well as the use of these antigens in an immunodiagnostic assay for the detection of antibodies againstin an isolated sample.

Soluble immunoreactive treponema pallidum TpN47 antigens

The present invention relates to fusion proteins suitable as test antigens in the detection of infections with pathogens, particularly of primary infections with pathogens. Further, the invention relates to methods for detecting and differentially determining antibodies, particularly IgM antibodies resulting from an infection with a pathogenic organism. Furthermore, test reagents for carrying out these methods are provided.

Detection of primary infections with pathogens

Disclosed are a recombinant DNA molecule encoding a fusion protein comprising a SlpA chaperone and a target polypeptide wherein human FK506 binding proteins (FKBPs) are excluded as target polypeptides, a corresponding expression vector encoding said fusion protein as well as host cells transformed with said expression vector. Also disclosed are a method for producing the fusion protein, a recombinantly produced fusion protein comprising a SlpA chaperone and a target polypeptide. A further aspect of the invention is the use of the recombinantly produced fusion protein, and a reagent kit containing a recombinantly produced fusion protein comprising a SlpA chaperone and a target polypeptide.

SlpA as a tool for recombinant protein and enzyme technology

The invention relates to soluble rubella E1 antigens and variants of these antigens. The antigens contain amino acids 201 to 432 or 169 to 432 and are lacking amino acids 453 to 481 as well as at least the amino acids 143 to 164. They further contain a region spanning two disulfide-bridges. The invention also relates to a recombinant DNA molecule encoding the rubella E1 antigens, the expression of rubella E1 antigens as chaperone fusion proteins and their use in a method of detecting antibodies against rubella in a sample.

Rubella E1 envelope protein variants and their use in detection of anti-rubella antibodies

The present invention relates to the cloning and expression of foreign protein or polypeptides in bacteria, such as. In particular, this invention relates to expression tools comprising a FKBP-type peptidyl prolyl isomerase selected from the group consisting of FkpA, SlyD, and trigger factor; methods of recombinant protein expression, the recombinant polypeptides thus obtained, as well as to the use of such polypeptides.

Methods for producing fusion polypeptides or enhancing expression of fusion polypeptides

The invention discloses the cloning, expression and uses of a chimeric fusion protein with superior chaperone and folding activities compared to the wild type chaperones. This invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule containing nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for an FK506 binding protein (FKBP) or an FK506-binding-protein-like domain (FKBP-like domain). In particular, this invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule containing nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for a human FKBP type peptidyl-prolyl-cis/trans isomerase (PPIase), methods of producing these chimeric fusion proteins and their uses as folding helpers in the production of other proteins and in the process of the production of vaccines or pharmaceuticals, and as folding helpers for performing immunoassays.

The invention discloses the cloning, expression and uses of a chimeric fusion protein with superior chaperone and folding activities compared to the wild type chaperones. This invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule comprising nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for an FK506 binding protein (FKBP) or an FK506-binding-protein-like domain (FKBP-like domain). In particular, this invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule comprising nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for a human FKBP type peptidyl-prolyl-cis/trans isomerase (PPIase), methods of producing these chimeric fusion proteins and their uses as folding helpers in the production of other proteins and in the process of the production of vaccines or pharmaceuticals, and as folding helpers for performing immunoassays.

A soluble rubella E1 antigen variant is disclosed that comprises amino acids 334-409 of the native rubella E1 peptide, but lacks the C-terminal end and at least the transmembrane region and the anchor segment as well as at least the amino acids 143 to 164. Also described is a recombinant DNA molecule encoding the rubella E1 antigen variants which are recombinantly expressed as a chaperone fusion protein, refolded into a soluble and immunoreactive conformation, and further used for the serological detection of anti-rubella antibodies. In addition, also disclosed is a method for the detection, determination and quantification of anti-rubella antibodies of IgG and/or IgM subclass in a sample wherein the rubella E1 antigen is used as a capture reagent and/or binding partner for the antibodies.

Soluble rubella E1 envelope protein variants

The present invention relates to the cloning and expression of foreign protein or polypeptides in bacteria, such asIn particular, this invention relates to expression tools comprising a FKBP-type peptidyl prolyl isomerase selected from the group consisting of FkpA, SlyD, and trigger factor; methods of recombinant protein expression, the recombinant polypeptides thus obtained, as well as to the use of such polypeptides.

Fusion polypeptides, vaccines and compositions of FKBP chaperones and target polypeptides

The present invention relates to the diagnosis of transmissible spongiform encephalopathies (TSEs). It especially teaches the production of a soluble prion protein (PrP)-chaperone complex and the advantageous use of such chaperone-PrP complex, especially in the detection of PrP in an immunoassay, as well as its use as and immunogen.

Complexes comprising a prion protein and a peptidyl prolyl isomerase chaperone, and method for producing and using them

The present invention relates to the diagnosis of Alzheimer's Disease. It especially teaches the production of a soluble A&#x3b2;-chaperone complex and the advantageous use of such chaperone-A&#x3b2; complex, especially in the detection of A&#x3b2; in an immunoassay, as well as its use as an immunogen.

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Uffing, Germany

Peter Schaarschmidt