Method of obtaining antibodies
First Claim
1. 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 NH2 terminus of the hemoglobin β
-chain;
(b) reacting the peptide of step (a) with glucose to obtain a glycosylated peptide having the formula
space="preserve" listing-type="equation">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.
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Accused Products
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 NH2 -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.
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Citations
6 Claims
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1. 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:
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(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 NH2 terminus of the hemoglobin β
-chain;(b) reacting the peptide of step (a) with glucose to obtain a glycosylated peptide having the formula
space="preserve" listing-type="equation">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. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification