Development of an enzyme-linked immunosorbent assay for human metallothionein-1 in plasma and urine.
The development of a sensitive enzyme-linked immunosorbent assay (ELISA) for human metallothionein-1 is reported. Metallothionein was purified from postmortem human liver and used to raise high-titer antibodies in rabbits. The assay was specific for human metallothionein-1 (MT-1), and there was no significant cross-reaction with human metallothionein-2. The detection limit (sensitivity) of the assay was 5 ng/ml, and the added MT-1 could be fully recovered from plasma and urine. The normal reference range for MT-1 was 32 +/- 16 ng/ml in plasma and 10 +/- 6 ng MT-1 per micromole of creatinine in random samples of urine. No significant differences were found between the values for males and females. The concentration of MT-1 was greatly increased between 24 and 48 hours after surgery, indicating that the protein behaves like an acute phase reactant in human subjects.
Akintola
D F
DF
Department of Chemical Pathology, Charing Cross and Westminster Medical School, London, United Kingdom.
Sampson
B
B
Fleck
A
A
D000209:Acute-Phase Proteins / Q000378:metabolism; D000210:Acute-Phase Reaction / Q000276:immunology / Q000378:metabolism; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000918:Antibody Specificity; D002850:Chromatography, Gel; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D008099:Liver / Q000737:chemistry; D008297:Male; D008668:Metallothionein / Q000097:blood* / Q000276:immunology / Q000652:urine*; D008875:Middle Aged; D011817:Rabbits; D012680:Sensitivity and Specificity
D000209:Acute-Phase Proteins; D000906:Antibodies; D008668:Metallothionein