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EP0133694A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Process for producing neuraminidase and method and reagent for the quantitative determination of a sialic acid-containing substance using neuraminidase

Assignee: KH Neochem Co Ltd
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Filing: 1984-08-03
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Granted: —
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Expires: 2004-08-03
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Abstract

Abstract Neuraminidase is produced by culturing a neuraminidase producing microorganism belonging to the genus Micromonospora. The neuraminidase is useful for the quantitative determination of a sialic acid-containing substance in a sample.

Inventors

  • 👤 Takayuki Uwajima
  • 👤 Kazuo Aisaka

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-03 12:30
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N1/00; C12N1/20; C12N1/205; C12N9/2402; C12Q; C12Q1/00; C12Q1/34; C12Y; C12Y302/00; C12Y302/01; C12Y302/01018; C12R; C12R2001/00; C12R2001/01; C12R2001/29; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/867; Y10S435/868

Claims Analysis

Total Claims: 16
Claim 1
Independent Claim 33 words
A process for producing neuraminidase which comprises culturing a neuraminidase producing microorganism belonging to the genus Micromonospora in a culture medium, accumulating neuraminidase in the culture liquor and thereafter recovering the neuraminidase therefrom.
Claim 2
Dependent Claim 33 words
A process for producing neuraminidase which comprises culturing a neuraminidase producing microorganism belonging to the genus Micromonospora in a culture medium, accumulating neuraminidase in the culture liquor and thereafter recovering the neuraminidase therefrom.
Claim 3
Dependent Claim 20 words
A process according to claim 1, wherein said microorganism belongs to the species Micromonospora viridifaciens, Micromonospora globosa or Micromonospora chalcea.
Claim 4
Dependent Claim 25 words
A process according to claim 2, wherein said microorganism is Micromonospora viridifaciens ATCC 31146, Micromonospora qlobosa NRRL B-2673 or Micromonos p ora chalcea ATCC 12452.
Claim 5
Dependent Claim 28 words
A process according to claim 1, wherein said culturing step is carried out at 25 to 35°C for 4 to 5 days at p H 6.0 to 7.0.
Claim 6
Independent Claim 109 words
A method for the quantitative determination of a sialic acid-containing substance in a sample which comprises: decomposing said sialic acid-containing substance in the sample with neuraminidase derived from a microorganism belonging to the genus Micromonos p ora to form N-acetylneuraminic acid; decomposing said N-acetylneuraminic acid with N -acetylneuraminic acid lyase to form pyruvic acid; reacting the formed pyruvic acid with NADH in the presence of lactic acid dehydrogenase and determining the degree of reduction in the amount of NADH; or reacting the formed pyruvic acid with pyruvate oxidase in the presence of peroxidase, thiamine pyrophosphate, 4-aminoantipyrine, phenol and magnesium salt and a colorimetrically determining the amount of pigment formed.
Claim 7
Independent Claim 109 words
A method for the quantitative determination of a sialic acid-containing substance in a sample which comprises: decomposing said sialic acid-containing substance in the sample with neuraminidase derived from a microorganism belonging to the genus Micromonos p ora to form N-acetylneuraminic acid; decomposing said N-acetylneuraminic acid with N -acetylneuraminic acid lyase to form pyruvic acid; reacting the formed pyruvic acid with NADH in the presence of lactic acid dehydrogenase and determining the degree of reduction in the amount of NADH; or reacting the formed pyruvic acid with pyruvate oxidase in the presence of peroxidase, thiamine pyrophosphate, 4-aminoantipyrine, phenol and magnesium salt and a colorimetrically determining the amount of pigment formed.
Claim 8
Dependent Claim 25 words
A method according to claim 5, wherein said neuraminidase is derived from a microorganism belonging to the species Micromonoposora viridifaciens, Micromonospora globosa or Micromonospora chalcea.
Claim 9
Dependent Claim 26 words
A method according to claim 5, wherein said neuraminidase is derived from Micromonospora viridifaciens ATC C 31146, Micromonospora globosa NRRL B-2673 or Micromonospora chalcea ATCC 12452.
Claim 10
Dependent Claim 25 words
A method according to claim 5, wherein said sialic acid-containing substance is selected from the group consisting of mucin, futuin, immunoglobulin, erythrocyte, serum or ganglioside.
Claim 11
Dependent Claim 22 words
A method according to claim 5, wherein said magnesium salt is MgSO 4 ·7H 2 O or MgCl 2 ·6H 2 O.
Claim 12
Dependent Claim 56 words
A reagent for the quantitative determination of a sialic acid-containing substance in sample which comprises: 0.5 to 2.0U of neuraminidase derived from a microorganism belonging to the genus Micromonospora; 1 to 10U of N-acetylneuraminic acid lyase; and a mixture of 0.3 to 0.9 pmole of NA D H and 2 to 5U of lactic acid dehydrogenase.
Claim 13
Dependent Claim 56 words
A reagent for the quantitative determination of a sialic acid-containing substance in sample which comprises: 0.5 to 2.0U of neuraminidase derived from a microorganism belonging to the genus Micromonospora; 1 to 10U of N-acetylneuraminic acid lyase; and a mixture of 0.3 to 0.9 pmole of NA D H and 2 to 5U of lactic acid dehydrogenase.
Claim 14
Dependent Claim 76 words
A reagent for the quantitative determination of a sialic acid-containing substance in a sample which comprises: 0.5 to 2.0U of neuraminidase derived from a microorganism belonging to the genus Micromonospora; 1 to 10U of N -acetylneuraminic acid lyase, and a mixture of 5 to 10U pyruvate oxidase, 50 to 100 U of peroxidase, 1 to 5 mg of thiamine pyrophosphate, 1 to 5 mg of a magnesium salt, 1 to 5 mg of 4-aminoantipyrine and phenol.
Claim 15
Dependent Claim 76 words
A reagent for the quantitative determination of a sialic acid-containing substance in a sample which comprises: 0.5 to 2.0U of neuraminidase derived from a microorganism belonging to the genus Micromonospora; 1 to 10U of N -acetylneuraminic acid lyase, and a mixture of 5 to 10U pyruvate oxidase, 50 to 100 U of peroxidase, 1 to 5 mg of thiamine pyrophosphate, 1 to 5 mg of a magnesium salt, 1 to 5 mg of 4-aminoantipyrine and phenol.
Claim 16
Dependent Claim 29 words
A reagent according to claim 11 wherein said magensium salt is selected from the group consisting of Mg SO 4 ·7H 2 O and MgCl 2 ·6H 2 O.

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