Roche Diagnostics Operations, Inc.

Roche Diagnostics Gmbh

Hoffmann-la Roche Inc.

Roche Diabetes Care, Inc.

Max-planck-gesellschaft Zur Forderung Der Wissenschaften E.v.

F. Hoffmann-la Roche Ag

Rainer Schmuck

Thomas Meier

Zhixin Shao

Peter Kratzsch

Carina Horn

Wolfgang-Reinhold Knappe

Dieter Heindl

Bernd Schneidinger

Joachim Hoenes

Mara Boenitz-Dulat

Daniela Beck

Christoph Seidel

Eva Hoess

Wilhelm Tischer

Detlef Thym

Klaus Reichert

Friedrich Popp

Gabriele Pestlin

Harald Weisser

Herbert Von Der Eltz

Rainer Mueller

Harald Sobek

Waltraud Ankenbauer

Johann-Peter Thalhofer

Alfred Engel

Rolf Nagel

Bruno Frey

Hartmut Duefel

Christine Markert-Hahn

Claudia Gaessler-Dietsche

Manfred Schmidt

Bernhard Suppmann

Elmar Weinhold

Nelli Steinke

Wolfgang Roedel

Rainer M&uuml;ller

Eberhard Russmann

Markus Pajatsch

Wolfgang Rödel

Kerstin Kr&uuml;ger

Claudia Gässler-Dietsche

Jessica Laggerbauer

Hartmut Düfel

Annette Deufel

Peter Pallua

Gisela Betzl

Janet Kenklies

Jessica Strey

Nadine Bucci

Joachim Hönes

Uwe Wehnes

Johnny Staepels

Iris Lein

Daniela Bunk

Methods for stabilizing an enzyme by storing the enzyme in the presence of a stabilized coenzyme are disclosed. In addition, an enzyme stabilized with a stabilized coenzyme as well as the use thereof in test elements for detecting analytes are also disclosed. Other aspects include unique compositions, methods, techniques, systems and devices involving enzyme stabilization.

Stabilization of enzymes with stable coenzymes

The present invention concerns a method for determining an analyte as well as a diagnostic element suitable therefore. In one particular form, a method for determining an analyte includes contacting a sample containing the analyte with a diagnostic element comprising a dry reagent layer. The dry reagent layer contains a mutated dehydrogenase which is specific for the analyte and an artificial coenzyme. The method also includes determining at least one of analyte presence and an amount of the analyte.

Fast reaction kinetics of enzymes having low activity in dry chemistry layers

The disclosure concerns the enzymatic synthesis of stable analogues of nicotinamide adenine dinucleotide NAD/NADH and nicotinamide adenine dinucleotide phosphate NADP/NADPH, the so-called 'carba-NADs', i.e. analogues of NAD/NADH or NADP/NADPH, respectively, comprising a carbacyclic sugar instead of ribose.

Enzymatic synthesis of carba-NAD

A method for detecting sequence specific methylation in a biomolecule, comprising: (a) contacting the biomolecule with an S-adenosyl-L-methionine-dependent methyltransferase in the presence of a detectable cofactor of said methyltransferase; and (b) detecting whether the recognition sequence of said methyltransferase has been modified with the cofactor or a derivative thereof, wherein modification of the recognition sequence of said methyltransferase is indicative of an absence of methylation at said recognition sequence. Also disclosed is a cofactor specific for S-adenosyl-L-methionine-dependent methyltransferases, wherein said cofactor is an N-adenosylaziridine derivative with a reporter group attached to the 6 or 7 position of the adenine ring or attached to the aziridine ring. A complex of the cofactor and a methyltransferase a composition comprising the cofactor or the complex and the use of the cofactor or the complex for detecting sequence-specific methylation in DNA molecules are also disclosed.

Sequence-specific detection of methylation in biomolecules

It was found that a fragment of nativeDNA polymerase (TaqWT) lacking 288 N-terminal amino acids (Taq&#x394;288) possesses an increased thermostability over TaqWT, Taq&#x394;279, and Taq&#x394;289. The present invention therefore provides Taq&#x394;288, recombinant expression vectors encoding the same or derivatives thereof, as well as purification protocols for Taq&#x394;288. The invention also encompasses kits containing Taq&#x394;288 as well as the use of Taq&#x394;288 and kits containing Taq&#x394;288. In addition, the invention encompasses methods for the sequencing a nucleic acid template and methods for amplifying a target nucleic acid.

Thermostable Taq polymerase fragment

The present invention relates to a mutant protein of PQQ-dependent s-GDH characterized in that in at least one of the positions 122 and 124 the amino acid lysine is present, wherein these positions correspond to the amino acid positions known from thes-GDH wild-type sequence (SEQ ID NO: 2), it also discloses genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.

Thermostable mutants of pyrroloquinoline quinone dependent glucose dehydrogenase

A mutant of PQQ-dependent soluble glucose dehydrogenase (s-GDH; EC 1.1.5.2) is provided with improved specificity for glucose as compared to maltose, having a substitution of threonine at position 348 by either glycine, alamine or serine, wherein said mutant additionally comprises, at least one mutation for improving the stability of the mutant and one or more mutation(s) for improving the affinity of the mutant to glucose, and/or one or more mutation(s) for further improving the specificity of the mutant for glucose as compared to maltose, and wherein position 348 correspond to the amino acid positions known from thes-GDH wild-type sequence. Also disclosed are genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.

Mutants of pyrroloquinoline quinone dependent soluble glucose dehydrogenase

The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH), to genes encoding mutated s-GDH, to mutant proteins of s-GDH with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugar, especially of glucose in a sample.

Forms of soluble pyrroloquinoline quinone-dependent glucose dehydrogenase

The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH) comprising an amino acid insertion between positions 428 and 429 as corresponding to the amino acid sequence known from, to genes encoding such variant s-GDH, to proteins of such s-GDH variants with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugars, especially of glucose in a sample.

Genetically engineered pyrroloquinoline quinone dependent glucose dehydrogenase comprising an amino acid insertion

The present invention relates to oxidoreductase apoenzyme variants which are enzymatically inactive but have coenzyme-binding properties. Further, the present invention relates to DNA sequences encoding these oxidoreductase apoenzyme variants, expression vectors containing such DNA sequences and the use of these oxidoreductase apoenzyme variants in diagnostic applications.

Inactivated enzyme variants and associated process and reagent system

A process is disclosed for the production of an antifusogenic peptide by producing a fusion peptide of a length of about 14 to 70 amino acids in a prokaryotic host cell, comprising the steps, under such conditions that inclusion bodies of said fusion peptide are formed, of: (a) expressing in said host cell a nucleic acid encoding said fusion peptide consisting of a first peptide which is an antifusogenic peptide of a length of about 10 to 50 amino acids and a second peptide of a length of about 4 to 30 amino acids, said first peptide being N-terminally linked to said second peptide; (b) cultivating said host cell to produce said inclusion bodies; and (c) recovering said antifusogenic peptide from said inclusion bodies, wherein said recovered antifusogenic peptide consists of said fusion peptide or a peptide comprising the antifusogenic peptide of about 10 to 50 amino acids and which is a fragment cleaved from said fusion peptide. Inclusion bodies of the peptides are disclosed. Also disclosed is a nucleic acid encoding the fusion peptide consisting of a first peptide which is an antifusogenic peptide selected from the group of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and said sequences further consisting of glycine at the C terminal end, N-terminally linked to the second peptide.

Nucleic acids encoding fusion peptides

The present invention relates to mutated genes encoding variants of an-type creatinase (EC 3.5.3.3, alternative name: creatine amidinohydrolase), to the creatinase variants encoded by these genes, and to different applications of these variants of creatinase, particularly to their use for determining the creatinine and/or creatine concentration in a sample.

Variants of Erwinia-type creatinase

A conjugate of a tissue non-specific alkaline phosphatase (tns-AP) and dextran which can be obtained by reacting unglycosylated tns-AP with activated dextran by incubation in aqueous solution, stopping the reaction and isolating the conjugate from the solution. The conjugate obtained in this manner is suitable as a standard for the determination of alkaline phosphatase.

Conjugate of a tissue non-specific alkaline phosphatase and dextran, process for its production and use thereof

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Benediktbeuern, Germany

Rainer Schmuck