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EP0346316A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Fusion protein and its use

Assignee: Kabigen AB
•
Filing: 1989-06-08
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Granted: —
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Expires: 2009-06-08
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Abstract

Abstract Fusion protein comprising a protein fragment carrying proteolytic activity and a CD4 binding glycosylated fragment originating from HIV; DNA-sequence encoding the fusion protein; antibody directed against the fusion protein; plasmid or vector carrying the DNA-sequence; host cell harbouring such plasmsid or vector; methods for determining HIV-antibodies and CD4; and method for the purification of the fusion protein.

Inventors

  • 👤 Magnus Gidlund
  • 👤 Mats Lake
  • 👤 Björn Löwenadler
  • 👤 Hans Wigzell

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-10-24 20:14
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; A; A61; A61P; A61P31/00; A61P31/04; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N9/6459; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069; C07K2319/00; C07K2319/35; C07K2319/70; C07K2319/74; C07K2319/75; C07K2319/90; C07K2319/91; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/16011; C12N2740/16111; C12N2740/16122

Claims Analysis

Total Claims: 25
Claim 1
Independent Claim 18 words
Fusion protein comprising a protein fragment carry­ing proteolytic activity and a CD4 binding glycosylated frag­ment originating from HIV.
Claim 2
Dependent Claim 18 words
Fusion protein comprising a protein fragment carry­ing proteolytic activity and a CD4 binding glycosylated frag­ment originating from HIV.
Claim 3
Dependent Claim 14 words
Fusion protein according to claim 1, wherein said protein fragment is a t-PA fragment.
Claim 4
Dependent Claim 23 words
Fusion protein according to claim 1 or 2, wherein said t-PA fragment comprises kringle 2 and the proteolytic portion of the t-PA molecule.
Claim 5
Dependent Claim 15 words
Fusion protein according to claim 1, 2 or 3, wherein said t-PA fragment is glycosylated.
Claim 6
Dependent Claim 15 words
Fusion protein according to claim 1, 2 or 3, wherein said t-PA fragment is non-glycosylated.
Claim 7
Dependent Claim 15 words
Fusion protein according to any preceding claim, wherein said CD4 binding fragment originates from HIV-envelo­pe.
Claim 8
Dependent Claim 12 words
Fusion protein according to claim 6, wherein said fragment originates from HIV-gp120.
Claim 9
Dependent Claim 11 words
DNA-sequence encoding the fusion protein of any of the preceding claims.
Claim 10
Dependent Claim 11 words
DNA-sequence according to claim 8, comprising the signal sequence of t-PA.
Claim 11
Dependent Claim 14 words
DNA-sequence according to claim 8 or 9, comprising also the leader sequence of t-PA.
Claim 12
Dependent Claim 22 words
Antibody directed against the fusion protein of any of claims 1 or 7 or against a CD4 binding fragment of said protein.
Claim 13
Dependent Claim 15 words
A plasmid or vector carrying the DNA sequence of any of claims 8 to 10.
Claim 14
Dependent Claim 12 words
A host cell harbouring the plasmid or vector accor­ding to claim 12.
Claim 15
Dependent Claim 12 words
A host cell according to claim 13 which is a euca­ryotic cell.
Claim 16
Dependent Claim 12 words
A host cell according to claim 14 which is a mamma­lian cell.
Claim 17
Independent Claim 97 words
A method for determining the presence of antibodies against HIV in a specimen, said antibodies reacting with re­gions within the CD4 binding fragment derived from the prima­ry sequence of HIV-envelope, comprising the steps: a) contacting said specimen with a fusion protein compri­sing a t-PA fragment carrying proteolytic activity, and a CD4 binding glycosylated fragment originating from HIV, said con­tact resulting in interaction between the antibodies of said specimen and said fusion protein; b) analyzing the product resulting from said interaction using the proteolytic activity of the product; and c) evaluating the contents of antibodies of said specimen.
Claim 18
Independent Claim 97 words
A method for determining the presence of antibodies against HIV in a specimen, said antibodies reacting with re­gions within the CD4 binding fragment derived from the prima­ry sequence of HIV-envelope, comprising the steps: a) contacting said specimen with a fusion protein compri­sing a t-PA fragment carrying proteolytic activity, and a CD4 binding glycosylated fragment originating from HIV, said con­tact resulting in interaction between the antibodies of said specimen and said fusion protein; b) analyzing the product resulting from said interaction using the proteolytic activity of the product; and c) evaluating the contents of antibodies of said specimen.
Claim 19
Independent Claim 87 words
A method for detecting the presence of CD4 in solu­tion or expressed on the surface of mammalian cells, compri­sing the steps: a) contacting a specimen containing such cells with a fu­sion protein comprising a t-PA fragment carrying proteolytic activity, and a CD4 binding glycosylated fragment originating from HIV, said contact resulting in interaction between the CD4 of said cells and said fusion protein; b) analyzing the product resulting from said interaction using the proteolytic activity of the product; and c) evaluating the CD4 contents of said cells.
Claim 20
Independent Claim 87 words
A method for detecting the presence of CD4 in solu­tion or expressed on the surface of mammalian cells, compri­sing the steps: a) contacting a specimen containing such cells with a fu­sion protein comprising a t-PA fragment carrying proteolytic activity, and a CD4 binding glycosylated fragment originating from HIV, said contact resulting in interaction between the CD4 of said cells and said fusion protein; b) analyzing the product resulting from said interaction using the proteolytic activity of the product; and c) evaluating the CD4 contents of said cells.
Claim 21
Dependent Claim 23 words
A method according to claim 16 or 17, wherein said t-PA fragment comprises kringle 2 and the proteolytic protein of the t-PA molecule.
Claim 22
Dependent Claim 17 words
A method according to claim 16, 17 or 18, wherein said CD4 binding fragment originates from HIV-envelope.
Claim 23
Dependent Claim 12 words
A method according to claim 19, wherein said frag­ment originates from HIV-gp120.
Claim 24
Dependent Claim 57 words
A method for the purification of the fusion protein according to any of claims 1 to 7, comprising the steps: a) immobilizing antibodies according to claim 9 to an in­soluble carrier; b) contacting a material containing said fusion protein with said immobilized antibodies resulting in interaction between same; and c) eluating the fusion protein from the carrier.
Claim 25
Dependent Claim 15 words
A method according to claim 21, wherein the elution is made by lowering the pH.

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