Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins

Eric E. Niederkofler, Kemmons A. Tubbs, Urban A. Kiernan, Dobrin Nedelkov, Randall W. Nelson

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

Novel mass spectrometric immunoassays (MSIAs) for the isolation and structural characterization of plasma apolipoprotein A-I (apoA-I), apoA-II, and apoE have been developed. The assays combine selective isolation of apolipoprotein species via affinity capture with mass-specific detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In application, plasma (from 50 μl of whole blood drawn from individuals, using finger lancet) was addressed with affinity pipette tips derivatized with antibodies toward the specific apolipoprotein. The time required for each assay was ∼15 min, less if assays on multiple individuals were performed in parallel. In a brief study of five individuals, several recently reported apoA-II variants were identified and observed consistently in all individuals. Additionally, the apoE phenotype of E3/E3 was observed in three of the individuals, and E2/E3 and E3/E4 observed in the remaining two individuals, the latter of whom suffers from Alzheimer's disease. Overall, the MSIA approach offers a rapid, sensitive, and highly accurate means of profiling apolipoproteins from small volumes of plasma.

Original languageEnglish (US)
Pages (from-to)630-639
Number of pages10
JournalJournal of Lipid Research
Volume44
Issue number3
DOIs
StatePublished - Mar 2003

Fingerprint

Apolipoproteins
Immunoassay
Apolipoprotein A-II
Assays
Apolipoproteins E
Plasmas
Plasma applications
Plasma Volume
Apolipoprotein A-I
Fingers
Ionization
Mass spectrometry
Desorption
Mass Spectrometry
Alzheimer Disease
Lasers
Blood
Phenotype
Antibodies

Keywords

  • Affinity capture
  • apoA-II
  • apoE
  • MALDI-TOF MS
  • Phenotype
  • Point mutations
  • Posttranslational modifications

ASJC Scopus subject areas

  • Endocrinology

Cite this

Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins. / Niederkofler, Eric E.; Tubbs, Kemmons A.; Kiernan, Urban A.; Nedelkov, Dobrin; Nelson, Randall W.

In: Journal of Lipid Research, Vol. 44, No. 3, 03.2003, p. 630-639.

Research output: Contribution to journalArticle

Niederkofler, Eric E. ; Tubbs, Kemmons A. ; Kiernan, Urban A. ; Nedelkov, Dobrin ; Nelson, Randall W. / Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins. In: Journal of Lipid Research. 2003 ; Vol. 44, No. 3. pp. 630-639.
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