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Measurement of Hemoglobin‐Acetaldehyde Adduct in Alcoholic Patients
Author(s) -
Lin Renee C.,
Shahidi Samin,
Kelly Timothy J.,
Lumeng Carey,
Lumeng Lawrence
Publication year - 1993
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.1993.tb00817.x
Subject(s) - acetaldehyde , keyhole limpet hemocyanin , hemoglobin , chemistry , adduct , alcohol , amino acid , in vivo , peptide , antibody , ethanol , lysine , biochemistry , medicine , biology , immunology , organic chemistry , microbiology and biotechnology
A sensitive and specific test for chronic alcohol abuse is useful in the diagnosis and management of alcoholic patients. Herein, we report the measurement of hemoglobin‐acetaldehyde adducts (Hb‐AA) in alcoholic patients by a sandwich ELISA using different antibodies. Keyhole limpet hemocyanin (KLH), a peptide consists of eight amino acid residues (8‐pep, V1 to K8) at the N‐terminus of βchain‐of human sickle‐cell Hb and a segment of HbA β‐chain consists of 11 amino acids rich in lysine or K (11‐pep, G56‐K66) were incubated with acetaldehyde and NaCNBH 3 to form protein‐AAs. 8‐Pep‐AA and 11‐pep‐AA were individually conjugated to unmodified KLH as the carrier. Anti‐protein‐AA lgGs were raised in rabbits using these three protein‐AA immunogens. When anti‐KLH‐AA IgG was‐used in ELISA, optical densities for alcoholic patients and controls were 0.311 ± 0.124 and 0.147 ± 0.042 (means ± SD, n = 40/group, p < 0.001), respectively. Using mean value ± 2 SD of controls as the cut‐off, sensitivities to detect alcoholic patients were 78, 75, and 43%, respectively, when anti‐KLH‐AA, anti‐11‐pep‐AA, and anti‐8‐ pep‐AA were used. Correlation among optical densities obtained from the first two lgGs was excellent ( R 2 = 0.905). We conclude that:(1) Hb‐AA has the potential of being a good marker for alcohol abuse, and (2) the site of Hb that is modified by acetaldehyde in vivo is‐ primarily located in a surface‐accessible domain near the center of the βchain of HbA where several lysine residues are clustered.