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Clinical Chemistry 47: 1249-1255, 2001;
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(Clinical Chemistry. 2001;47:1249-1255.)
© 2001 American Association for Clinical Chemistry, Inc.


Articles

Immunochemical Assay of Hemoglobin with N{epsilon}-(Carboxymethyl)lysine at Lysine 66 of the ß Chain

Hisahiko Iwamoto1a, Yoshihiro Motomiya2, Keisuke Miura1, Masayo Morisawa2, Yoshimichi Yoshimura1 and Ikuro Maruyama3

1 R&D for Diagnostics, A&T Corporation, 2023-1, Endo, Fujisawa-City, Kanagawa 252-0816, Japan.
2 Suiyukai Clinic, 676-1, Kuzumoto-chou, Kashihara-City, Nara 634-0007, Japan.
3 Department of Laboratory Medicine, Kagoshima University School of Medicine, Kagoshima 890-0075, Japan.


aAuthor for correspondence. Fax 81-466-86-8676; e-mail iwamoto{at}alice.aandt.co.jp.

Background: N{epsilon}-(Carboxymethyl)lysine (CML), a well-characterized and major advanced glycation end product structure, is produced via a Maillard reaction by nonenzymatic glycation and/or oxidation. Although few of the carboxymethylation sites of lysine residues on proteins have been identified, it is known that the possible lysine glycation site in hemoglobin (Hb) is Lys-66 on the ß chain. We aimed to develop an assay for the Hb with a CML (CML-Hb) site specific to Lys-66 on the Hb ß chain and to determine whether the lysine residue at that site is carboxymethylated.

Methods: Ala-His-Gly-Lys-Lys(CM)-Val-Leu-Gly-Ala-Phe-Ser-Cys, the peptide derived from the ß chain of human Hb, was synthesized as an immunogen, and a monoclonal antibody against the peptide was prepared. A latex immunoassay method was established using the antibody on an automatic analyzer. In this study, 20 samples from healthy subjects and 80 samples from nondiabetic patients undergoing hemodialysis (HD) were analyzed.

Results: The latex immunoassay method using the antibody correlated significantly with the ELISA method using the antibody (r = 0.95; P <0.001). Between healthy subjects (n = 20) and nondiabetic HD patients (n = 80), a significant difference was seen in circulating CML-Hb (525 ± 76 vs 778 ± 137 pmol CML/mg of Hb; P <0.0001).

Conclusion: The latex method for the CML-Hb site specific to Lys-66 on the ß chain can measure large numbers of samples on an automatic analyzer.




The following articles in journals at HighWire Press have cited this article:


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Clin. Chem.Home page
U. Friess, A. Beck, E. Kohne, R. Lehmann, S. Koch, H.-U. Haring, R.-M. Schmuelling, and E. Schleicher
Novel Hemoglobin Variant [{beta}66(E10) Lys->Asn], with Decreased Oxygen Affinity, Causes Falsely Low Hemoglobin A1c Values by HPLC
Clin. Chem., August 1, 2003; 49(8): 1412 - 1415.
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