Boehringer Mannheim Gmbh

Roche Diagnostics Gmbh

Walter Schneider

Albert Roder

Sigmar Klose

Ulfert Deneke

Gunter Weimann

Klaus Beaucamp

Klaus Muhlegger

Hans Mollering

Josef Burg

Peter Stahl

Gerhard Michal

Hans-Otto Beutler

Karl-Heinz Sellinger

Ingeborg Gutmann

Rolf Laube

Ingrid Schutte

Werner F&uuml;rst

Alexander Wrba

A process for producing erythropoietin which is free of foreign animal proteins except for the proteins of the host cell, wherein DNA coding for EPO is expressed in a eukaryotic host cell and the host cell is cultured in a medium free of natural mammalian proteins. The erythropoietin is chromatographically purified using dye affinity chromatography, chromatography on hydroxyapatite, reversed phase chromatography, and anion exchange chromatography. The resulting preparation contains less than 100 ppm of proteins derived from the host cell, and less than 10 pg of host cell DNA per 83 &mgr;g erythropoietin.

Process for producing erythropoietin containing no animal proteins

The present invention provides a process for the enzymatic determination of sulphite, wherein the sulphite is oxidized in a buffered solution with sulphite oxidase to give sulphate, the hydrogen peroxide thereby formed being reduced in the presence of NAD and NADH with the enzyme NADH peroxidase and the consumption of NADH being determined optically. The present invention also provides a reagent for the enzymatic determination of sulphite, wherein it contains sulphite oxidase, NADH, NAD, NADH peroxidase and buffer.

Process and reagent for the enzymatic determination of sulphite

A method for the determination of triglycerides by ester cleavage utilizing lipase and optionally esterase with the formation of fatty acids and glycerol, phosphorylation of the glycerol with adnosine triphosphate in the presence of glycerol kinase with the formation of glycerol-1-phosphate and adenosine diphosphate and determination of one of the latter two products. The glycerol kinase used is from Bacillus stearothermophilis and acts in combination with at least one activator selected from the group consisting of detergents, phenol derivatives and aniline derivative.

Method and reagent for the determination of triglycerides

In a method for the determination of glutamate-oxalacetate transaminase or glutamate-pyruvate transaminase by the reaction of oxalacetate or pyruvate with glutamate, with formation of .alpha.-ketoglutarate in a buffered solution, the .alpha.-ketoglutarate formed is reacted with .gamma.-aminobutyrate in the presence of .gamma.-aminobutyrate transaminase with formation of succinate semialdehyde, NADP is reduced with the latter in the presence of succinate semialdehyde dehydrogenase to give NADPH, and the latter is measured either directly or after conversion with a tetrazolium salt and an electron carrier such as diaphorase, phenantroline methosulphate or phenazine methosulphate to a formazane dye. A reagent suited for use with this method contains .gamma.-aminobutyrate transaminase, succinate semialdehyde dehydrogenase, .gamma.-aminobutyrate, glutamate and buffer, as well as either oxalacetate or pyruvate, and optionally a tetrazolium salt, an electron carrier and a surface-active agent.

Method and reagent for the determination of glutamate-oxalacetate

Formate, or compounds convertible into formate, is determined by reacting same in the presence of formate dehydrogenase (FDH) and of a hydrogen acceptor, e.g., NAD, wherein the formate dehydrogenase is from Candida boidinii DSM 941, to result in stoichiometric reactions. The process can be used for the removal of formate from solutions containing same.

Process for determining formate and reagent therefor

Stabilized urease compositions are provided comprising urease and, as a stabilizing agent, a mixture of glutathione, ethylenediamine-tetraacetic acid, and citrate, preferably in a weight ratio of 1:0.5-5:0.5-3:0.5-3.

Stabilized urease composition

NADP-dependent dehydrogenases are separated and purified by subjecting mixtures containing it to affinity chromatography using as the affinity chromatography material a nucleotide with a phosphate group in the 2'- or 3'-position, bound to a solid carrier.

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Weilheim, Germany

Walter Schneider