US4478744A
Expired / Lapsed
US
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Method of obtaining antibodies
Assignee: Roche Diagnostics GmbH
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Filing: 1982-01-25
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Granted: 1984-10-23
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Expires: 2002-01-25
Abstract
Abstract Antibodies specific to glycosylated hemoglobin and substantially free of cross-reactivity with nonglycosylated forms of hemoglobin are obtained by administering a synthetic antigen to a host animal. The antigen comprises a glycosylated synthetic peptide having an amino acid sequence corresponding to that of the NH 2 -terminus of the β-chain of hemoglobin. The peptide comprises a sequence of between about four and ten, and preferably seven, amino acids, and is conjugated with glucose and a protein. The protein preferably comprises an immunoglobulin which is foreign to the host animal.
Inventors
- 👤 Louis M. Mezei
- 👤 Jung S. Chen
- 👤 John J. Huang
- 👤 Robert E. Lovins
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 10:20
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/72; G01N33/721; G01N33/723; C; C07; C07K; C07K14/00; C07K14/795; C07K14/805; Y; Y10; Y10S; Y10S530/00; Y10S530/806
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
128 words
A method of preparing an antigen useful in obtaining antibodies against HbAlc, which antibodies are substantially free of cross-reactivity to nonglycosylated hemoglobin, said method comprising the steps of: (a) synthesizing a hapten consisting of a peptide having the formula Val--His--Leu--Y wherein Y is chosen from the group consisting of Thr--, Thr--Pro--, Thr--Pro--Glu--, Thr--Pro--Glu--Glu--, Thr--Pro--Glu--Glu--Lys--, Thr--Pro--Glu--Glu--Lys--Ser--, and Thr--Pro--Glu--Glu--Lys--Ser--Ala--, said hapten thereby having an amino acid sequence corresponding to the NH 2 terminus of the hemoglobin β-chain; (b) reacting the peptide of step (a) with glucose to obtain a glycosylated peptide having the formula glucose--Val--His--Leu--Y; and (c) conjugating the glycosylated peptide of step (b) with a protein to form an antigen suitable for injection into a host animal whose metabolism does not naturally produce HbAlC for production of antibodies thereto.
Claim 2
Independent Claim
128 words
A method of preparing an antigen useful in obtaining antibodies against HbAlc, which antibodies are substantially free of cross-reactivity to nonglycosylated hemoglobin, said method comprising the steps of: (a) synthesizing a hapten consisting of a peptide having the formula Val--His--Leu--Y wherein Y is chosen from the group consisting of Thr--, Thr--Pro--, Thr--Pro--Glu--, Thr--Pro--Glu--Glu--, Thr--Pro--Glu--Glu--Lys--, Thr--Pro--Glu--Glu--Lys--Ser--, and Thr--Pro--Glu--Glu--Lys--Ser--Ala--, said hapten thereby having an amino acid sequence corresponding to the NH 2 terminus of the hemoglobin β-chain; (b) reacting the peptide of step (a) with glucose to obtain a glycosylated peptide having the formula glucose--Val--His--Leu--Y; and (c) conjugating the glycosylated peptide of step (b) with a protein to form an antigen suitable for injection into a host animal whose metabolism does not naturally produce HbAlC for production of antibodies thereto.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein said protein is an immunoglobulin.
Claim 4
Dependent Claim
10 words
The method of claim 2 wherein said protein comprises IgG.
Claim 5
Dependent Claim
14 words
The method of claim 1 wherein said peptide is covalently linked to said protein.
Claim 6
Dependent Claim
17 words
The method of claim 4 wherein said peptide is conjugated to said protein through a carbodiimide linkage.
Claim 7
Dependent Claim
29 words
The method of claim 2 wherein said peptide comprises d-glucose--Val--His--Leu--Thr--Pro--Glu--Glu--, said protein is bovine IgG, and said peptide is covalently linked to said bovine IgG through a carbodiimide linkage.
References Cited (Backward Citations)
No citation records indexed.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-10-23 | 1988-04-23 | 1988-10-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-10-23 | 1992-04-23 | 1992-10-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-10-23 | 1996-04-23 | 1996-10-23 | $7700.00 | Window Closed / Lapsed |