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Journal of Lipid Research, Vol. 44, 630-639, March 2003
Copyright © 2003 by Lipid Research, Inc.
Methods |
Intrinsic Bioprobes, Inc., 625 South Smith Road, Suite 22, Tempe, AZ 85281
1 To whom correspondence should be addressed. e-mail: rnelson{at}intrinsicbio.com
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.
Abbreviations: CDI, carbonyldiimidazole; CMD, caboxymethyl dextran; HBS, Hepes-buffered saline; MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; MSIA, mass spectrometric immunoassay; PTM, posttranslational modifications; SNP, single-nucleotide polymorphism; TCEP, tris-2-carboxyethyl phosphine hydrochloride
Supplementary key words MALDI-TOF MS • affinity capture • apoE • apoA-II • posttranslational modifications • point mutations • phenotype
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