US4735896A
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US
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Synthetic peptide and process of using same for the detection and diagnosis of AIDS and pre-AIDS conditions
Assignee: United Biomedical Inc
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Filing: 1986-04-02
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Granted: 1988-04-05
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Expires: 2006-04-02
Abstract
Abstract The present invention relates to a method for the detection and diagnosis of AIDS (acquired immune deficiency syndrome) ARC (AIDS Related Complex) and pre-AIDS conditions in body fluids by the use of a chemically synthesized peptide. The peptide has an amino acid sequence corresponding to a segment of the envelope protein, p41, of HTLV-III and has been found to be highly immunoreactive with antibodies in sera of patients with AIDS, ARC and pre-AIDS conditions. More specifically, the present invention is directed to the use of a chemically synthesized peptide containing therein a segment of about twenty-one (21) amino acids, or their analogues, in a prescribed sequence for the detection of antibodies to the HTLV-III virus in human body fluids of AIDS, ARC or pre-AIDS patients. The detection method includes an enzyme-linked immunosorbent assay (ELISA), an immunoradiometric assay (IRMA), and other forms of immuno assay procedures such as enzyme immuno blotting assay on nitrocellulose paper and hemagglutination assay using the peptide as the antigen. The preferred detection method is ELISA. The present invention also relates to a vaccine and a method for generating antibodies to HTLV-III in healthy mammals, including humans by the use of the chemically synthesized 21mer peptide or its analogues.
Inventors
- 👤 Chang Y. Wang
- 👤 James J. G. Wang
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 10:15
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; C07K16/00; C07K16/08; C07K16/10; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; A; A61; A61K; A61K39/00; C12; C12N; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/16011; C12N2740/16111; C12N2740/16122; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/81; Y10S435/974; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/811; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815; Y10S930/00; Y10S930/01; Y10S930/22; Y10S930/221
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
97 words
Peptide composition having specific immunoreactivity to antibodies to HTLV-III selected from the group consisting of peptide containing from twelve to twenty-five amino acids, the sequence of which corresponds to a part or the entirety of the following sequence: Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser; analogues thereof wherein the amino acids in the sequence are substituted, deleted or added as long as the immunoreactivity to antibodies to HTLV-III derived from the three dimensional conformation of the sequence is preserved; and conjugates of the peptides or analogues thereof, wherein: Ala=alanine, Arg=arginine, Cys=cysteine Gln=glutamine, Glu=glutamic acid, Gly=glycine, Ile=isoleucine, Leu=leucine, Lys=lysine, Ser=serine, Trp=tryptophan, Tyr=tyrosine, and Val=valine.
Claim 2
Dependent Claim
97 words
Peptide composition having specific immunoreactivity to antibodies to HTLV-III selected from the group consisting of peptide containing from twelve to twenty-five amino acids, the sequence of which corresponds to a part or the entirety of the following sequence: Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser; analogues thereof wherein the amino acids in the sequence are substituted, deleted or added as long as the immunoreactivity to antibodies to HTLV-III derived from the three dimensional conformation of the sequence is preserved; and conjugates of the peptides or analogues thereof, wherein: Ala=alanine, Arg=arginine, Cys=cysteine Gln=glutamine, Glu=glutamic acid, Gly=glycine, Ile=isoleucine, Leu=leucine, Lys=lysine, Ser=serine, Trp=tryptophan, Tyr=tyrosine, and Val=valine.
Claim 3
Dependent Claim
12 words
A peptide composition according to claim 1 wherein the peptide is: Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser.
Claim 4
Dependent Claim
81 words
An immunoassay method for the detection of antibodies to HTLV-III and diagnosis of AIDS, ARC or pre-AIDS conditions comprising: (a) coating a solid support with an effective amount of the peptide composition according to claim 1 as an antigen; b. adding a test sera diluted with a buffer wherein the antibodies to HTLV-III in the test sera form a peptide-antibody complex with the peptide composition; c. incubating the mixture at room temperature; and d. detecting the presence of the peptide-antibody complex.
Claim 5
Dependent Claim
23 words
An immunoassay method according to claim 3 wherein the solid support is coated with peptide having twenty-one amino acids in a sequence: Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser.
Claim 6
Dependent Claim
33 words
An immunoassay method according to claim 3 wherein step d. comprises: introducing a second known antibody labelled with an enzyme and a substrate which reacts with the enzyme to a form colored product.
Claim 7
Dependent Claim
21 words
An immunoassay method according to claim 2 wherein step d. comprises: introducing a second known antibody labelled with a radioactive element.
Claim 8
Dependent Claim
16 words
An immunoassay method according to claim 3 wherein the peptide-antibody complex is detectable as an agglutination.
Claim 9
Dependent Claim
38 words
An immunoassay method according to claim 5 wherein for each assay procedure, the amount of the peptide composition is about 0.1 ug to about 20 ug of the peptide in a buffer of about pH 7 to 10.
Claim 10
Dependent Claim
34 words
An immunoassay method according to claim 5 wherein for each assay procedure, the amount of the peptide composition is about 1 ug of the peptide in a buffer of about pH 7 to 10.
Claim 11
Dependent Claim
38 words
An immunoassay method according to claim 6 wherein for each assay procedure, the amount of the peptide composition is about 0.1 ug to about 20 ug of the peptide in a buffer of about pH 7 to 10.
Claim 12
Dependent Claim
34 words
An immunoassay method according to claim 6 wherein for each assay procedure, the amount of the peptide composition is about 1 ug of the peptide in a buffer of about pH 7 to 10.
Claim 13
Dependent Claim
70 words
A test kit for the detection of antibodies to HTLV-III and the diagnosis of AIDS, ARC and pre-AIDS conditions comprising: a. a solid support; b. coating onto the solid support, an immunoadsorbent comprising a peptide composition according to claim 1 c. a sample of normal serum as negative control; d. a sample of serum containing antibodies to HTLV-III as positive control; and e. a buffer for diluting the serum samples.
Claim 14
Dependent Claim
21 words
A test kit according to claim 12 wherein the immunoadsorbent is a peptide of twenty-one amino acids with a sequence: Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser.
Claim 15
Dependent Claim
50 words
A test kit according to claim 12 further comprising: f. an enzyme labelled second antibody for binding with the peptide-antibody complex; g. a substrate which reacts with the enzyme labelled antibody to form a colored product; and h. a second buffer solution for stopping the development of the colored product.
Claim 16
Dependent Claim
27 words
A test kit according to claim 12 further comprising: f. a radioactive element labelled second antibody for binding with the peptide-antibody complex to detect the peptide-antibody complex.
Claim 17
Dependent Claim
56 words
A peptide which is specifically immunoreactive with antibodies to HTLV-III with not more than twenty-five amino acids in a sequence having as a segment thereof selected from the group consisting of: (a) -Trp-Gly-Cys-Ser; (b) -Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile; and (c) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp- wherein Trp=tryptophan, Gly=glycine, Cys=cysteine, Ser=serine, Asp=Aspartic acid, Glu=glutamine, Leu=leucine, Ile=isoleucine, Ala=alanine, Val=valine, Glu=glutamic acid, Arg=arginine, Tyr=Tyrosine and Lys=lysine.
Claim 18
Dependent Claim
56 words
A peptide which is specifically immunoreactive with antibodies to HTLV-III with not more than twenty-five amino acids in a sequence having as a segment thereof selected from the group consisting of: (a) -Trp-Gly-Cys-Ser; (b) -Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile; and (c) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp- wherein Trp=tryptophan, Gly=glycine, Cys=cysteine, Ser=serine, Asp=Aspartic acid, Glu=glutamine, Leu=leucine, Ile=isoleucine, Ala=alanine, Val=valine, Glu=glutamic acid, Arg=arginine, Tyr=Tyrosine and Lys=lysine.
Claim 19
Dependent Claim
16 words
A peptide composition according to claim 16 wherein the peptide comprises at least fifteen amino acids.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1991-04-05 | 1991-10-05 | 1992-04-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-04-05 | 1995-10-05 | 1996-04-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-04-05 | 1999-10-05 | 2000-04-05 | $7700.00 | Window Closed / Lapsed |