Enzymatic assay of homocysteine on microtiter plates or a TECAN analyzer using crude lysate containing recombinant methionine γ-lyase

Err Cheng Chan, Pi Yueh Chang, Tsu Lan Wu, James T. Wu*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

23 Scopus citations

Abstract

An enzymatic assay for plasma homocysteine was developed that uses a crude lysate of E. coli containing the recombinant enzyme, methionine γ-lyase. The assay uses a commercially available fluorophore and 96-well microtiter plates; it can be performed manually or with the TECAN automated analyzer. The CVs for within-run and between-run precision are <10%. Close correlation (r >0.9) was obtained between results by this enzymatic method vs a reference HPLC procedure. In a Chinese population, the concentration of plasma total homocysteine was found to be gender- and age-dependent. Mean concentrations of plasma total homocysteine increased with age and were higher in men than women. Serum homocysteine concentrations did not differ significantly from those in plasma, provided the whole blood specimens were kept at 4°C for 2 hr, or at room temperature for <45 min, between venepuncture and centrifugation.

Original languageEnglish
Pages (from-to)155-160
Number of pages6
JournalAnnals of Clinical and Laboratory Science
Volume35
Issue number2
StatePublished - 03 2005

Keywords

  • Enzymatic assay
  • Homocysteine
  • Methionine γ-lyase
  • Microtiter plate
  • TECAN analyzer

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