Genotyping of apolipoprotein E by matrixassisted laser desorption/ionization time-of-flight mass spectrometry

Jannavi R. Srinivasan, Maureen T. Kachman, Anthony A. Killeen, Nahida Akel, David Siemieniak, David M. Lubman

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

The genotyping of the various isoforms of Apolipoprotein E (apo E) has been performed using matrixassisted laser desorption/ionization (MALDI-MS). The polymerase chain reaction was used to amplify the specific apo E gene sequence followed by digestion with Cfo I (Clostridium formicoaceticum), for generating restriction fragments for rapid and accurate mass analysis. An exonuclease I digestion step was introduced to remove the unused primers after PCR, which can otherwise interfere in the mass spectral analysis. By replacing the gel electrophoresis detection step with MALDI-MS, restriction isotyping of the apo E gene was achieved. Genotyping of an unknown sample obtained from an independent diagnostic laboratory demonstrated the validity of the MALDI-MS method for the routine clinical analysis of apo E.

Original languageEnglish (US)
Pages (from-to)1045-1050
Number of pages6
JournalRapid Communications in Mass Spectrometry
Volume12
Issue number16
DOIs
StatePublished - 1998

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