EP0138704A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Hepatitis A-subunit antigen
Assignee: Merck and Co Inc
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Filing: 1984-10-09
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Granted: —
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Expires: 2004-10-09
Abstract
Abstract @ A surface structural protein of Hepatitis A Virus (HAV) has been isolated and characterized from virus grown in tissue culture. This 33,00 dalton viral protein (VP-1) reacts with immune HAV sera and monoclonal antibodies that neutralize HAV infectivity. The VP-1 is usable for the preparation of a polypeptide subunit vaccine for HAV. Hybridoma cells were made which produced monoclonal antibodies to HAV or VP-1. These monoclonal antibodies were found to neutralize the infectivity of HAV and to compete with polyclonal antibody derived from human HAV immune sera. The monoclonal antibodies are useful for the neutralization of infectious HAV, the detection of antibodies to neutralizing sites on HAV, and the diagnoses of HAV disease in humans and other susceptible hosts.
Inventors
- 👤 Joseph V. Hughes
- 👤 Edward M. Scolnick
- 👤 Joanne E. Tomassini
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-10 00:41
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; C07K16/00; C07K16/08; C07K16/10; C07K16/106; A; A61; A61K; A61K38/00; A61K39/00; C12; C12N; C12N2770/00; C12N2770/00011; C12N2770/32011; C12N2770/32411; C12N2770/32422; C12N2770/32434; Y; Y02; Y02A; Y02A50/00; Y02A50/30; Y10; Y10S; Y10S530/00; Y10S530/806; Y10S530/82; Y10S530/826; Y10S930/00; Y10S930/01; Y10S930/22; Y10S930/223
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
68 words
A hepatitis A virus surface protein having a molecular weight of about 33,000 daltons, which protein reacts with neutralizing antibodies to hepatitis A virus and which contains after acid hydrolysis and gel electrophoresis the following amino acids in approximate order of decreasing percent by weight: glycine, serine, glutamic acid and/or glutamine, aspartic acid and/or asparagine, alanine, threonine, leucine, valine, lysine, proline, phenylalanine, arginine, isoleucine, tyrosine, histidine and methionine.
Claim 2
Dependent Claim
68 words
A hepatitis A virus surface protein having a molecular weight of about 33,000 daltons, which protein reacts with neutralizing antibodies to hepatitis A virus and which contains after acid hydrolysis and gel electrophoresis the following amino acids in approximate order of decreasing percent by weight: glycine, serine, glutamic acid and/or glutamine, aspartic acid and/or asparagine, alanine, threonine, leucine, valine, lysine, proline, phenylalanine, arginine, isoleucine, tyrosine, histidine and methionine.
Claim 3
Dependent Claim
148 words
A hepatitis A virus surface protein according to Claim 1 wherein after acid hydolysis and gel electrophoresis the amino acids are present in approximately the following percentages by weight: glycine, from about 21% to about 30%, serine, from about 9% to about 11%, glutamic acid and/or glutamine, from about 8% to about 10.5%, aspartic acid and/or asparagine, from about 7.2% to about 10% alanine, from about 6% to about 7.3%, threonine, from about 4.9% to about 8.2%, leucine, from about 4.9% to about 7.2%, valine, from about 4% to about 5.4%, lysine, from about 3.7% to about 4.3%, proline, from about 3.2% to about 5.3%, phenylalanine, from about 3% to about 4.5%, arginine, from about 2% to about 4.1%, isoleucine, from about 2% to about 3.6%, tyrosine, from about 2.0% to about 3.0%, histidine, from about 1.3% to about 1.9%, and methionine, from about 0.3% to about 0.9%.
Claim 4
Dependent Claim
36 words
A hepatitis A virus surface protein according to Claim 1 containing one or more of the following internal amino acid sequences from the direction of the amino terminus to the carboxy terminus of the peptide: Sequence
Claim 5
Dependent Claim
48 words
A method for isolating the surface protein of Claim 1 comprising: a) solubilizing the intact hepatitis A virus in a solution containing an anionic surfactant and a reducing agent; b) separating the viral proteins; and c) selecting the viral protein having a molecular weight of about 33,000 daltons.
Claim 6
Dependent Claim
14 words
A method according to Claim 4 wherein the anionic surfactant is sodium dodecyl sulfate.
Claim 7
Dependent Claim
12 words
A method according to Claim 4 wherein the reducing agent is 2-mercaptoethanol.
Claim 8
Dependent Claim
18 words
A method according to Claim 4 wherein the separating of the viral proteins is effected by gel electrophoresis.
Claim 9
Dependent Claim
21 words
A hybridoma cell made by fusion of a mammalian spleen cell immunized with hepatitis A virus to a mammalian myeloma cell.
Claim 10
Dependent Claim
21 words
A hybridoma cell made by fusion of a mammalian spleen cell immunized with hepatitis A virus to a mammalian myeloma cell.
Claim 11
Dependent Claim
28 words
A hybridoma cell according to Claim 8 wherein the immunized spleen cell is obtained from a mammal injected in vivo with hepatitis A virus or protein subunits thereof.
Claim 12
Dependent Claim
30 words
A hybridoma cell according to Claim 8 wherein the immunized spleen cell is obtained by in vitro sensitization of mammalian spleen cells with hepatitis A virus or protein subunits thereof.
Claim 13
Dependent Claim
14 words
A hybridoma cell according to Claim 8 wherein the mammalian cells are from mice.
Claim 14
Dependent Claim
24 words
A substantially, pure monoclonal antibody to hepatitis A virus having a molecular weight of about 150,000 daltons, and being subtype IgG-1 or subtype IgG-2a.
Claim 15
Dependent Claim
24 words
A substantially, pure monoclonal antibody to hepatitis A virus having a molecular weight of about 150,000 daltons, and being subtype IgG-1 or subtype IgG-2a.
Claim 16
Dependent Claim
15 words
A monoclonal antibody to hepatitis A virus produced by the hybridoma cell of Claim 8.
Claim 17
Dependent Claim
32 words
A diagnostic reagent comprising a surface having adhered thereto a inner layer of monoclonal antibody to hepatitis A virus, and an outer layer of hepatitis A virus adhered to the inner layer.
Claim 18
Dependent Claim
32 words
A diagnostic reagent comprising a surface having adhered thereto a inner layer of monoclonal antibody to hepatitis A virus, and an outer layer of hepatitis A virus adhered to the inner layer.
Claim 19
Dependent Claim
24 words
A diagnostic reagent according to Claim 14 wherein the inner layer of monoclonal antibody is adhered to an innermost layer of goat antimouse antibody.
Claim 20
Dependent Claim
10 words
16 . The 1125 labelled monoclonal antibody of Claim 12.
Claim 21
Dependent Claim
30 words
A competitive assay for determining HAV neutralizing antibody titer of a sample which comprises adding I 125 labelled monoclonal HAV or VP-1 antibody to the diagnostic reagent of Claim 14.
Claim 22
Dependent Claim
29 words
A competitive assay for determining HAV neutralizing antibody titer of a sample which comprises adding 1125 labelled monoclonal HAV or VP-1 antibody to the diagnostic reagent of Claim 15.
Claim 23
Dependent Claim
30 words
The method of inducing neutralizing antibodies against hepatitis A virus or protein subunits thereof in mammals by administering an effective amount to said mammals of the protein of Claim 3.
Claim 24
Dependent Claim
17 words
A therapeutic composition comprising a pharmaceutical carrier containing an effective amount of the protein of Claim 3.
Claim 25
Dependent Claim
25 words
A hepatitis A surface antigen protein according to Claim 1 which has the sequence wherein X is unknown and A is either Gln or Glu.
References Cited (Backward Citations)
No citation records indexed.
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