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A more recent version of this article appeared on March 1, 2003

Papers In Press, published online ahead of print December 1, 2002
J. Lipid Res., doi:10.1194/jlr.D200034-JLR200
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Submitted on September 4, 2002
Revised on November 4, 2002
Accepted on November 26, 2002

Novel Mass Spectrometric Immunoassays for the Rapid Structural Characterization of Plasma Apolipoproteins

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

Intrinsic Bioprobes Inc., Tempe, AR 85281

Corresponding Author: rnelson{at}intrinsicbio.com

Novel mass spectrometric immunoassays for the isolation and structural characterization of plasma apolipoproteins A-I, A-II and E 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 (MALDI-TOF MS). In application, plasma (from 50 mu L 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 minutes, less if assays on multiple individuals were performed in parallel. In a brief study of five individuals, several as yet un-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 which suffers from Alzheimer’s disease. Overall, the mass spectrometric immunoassay approach offers a rapid, sensitive and highly accurate means of profiling apolipoproteins from small volumes of plasma.


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