ProteinChip Technology: A New and Facile Method for the Identification and Measurement of High-Density Lipoproteins apoA-I and apoA-II and Their Glycosylated Products in Patients with Diabetes and Cardiovascular Disease

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Abstract
This paper describes a ProteinChip technology for the identification and quantification of apolipoprotein profiles in crude biological samples. Expression levels of apoA-I and apoA-II and their glycosylated products were accomplished using single 1 μL plasma samples. In the present studies, strong anionic and weak cationic exchanger ProteinChips (SAX2 and WCX2 chip surfaces) were tested, and the WCX2 chip was found to be selective for specific apolipoproteins. Using the WCX2 chip and analysis via surface-enhanced laser desorption ionization mass spectrometry (SELDI-MS), apoA-I and apoA-II were separated as sharp peaks at 28 and 17 kD and did not overlap with other serum protein peaks. Since these assays can be completed on a large number of clinical samples in approximately 1 h, further development of this technique will facilitate both epidemiological studies and therapeutic trials in assessing the role of the apolipoproteins and their glycosylated products in atherosclerosis. Keywords: diabetes • cardiovascular disease • surface-enhanced laser desorption/ionization (SELDI) • time-of-flight mass spectrometry (TOF-MS) • matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) • HDL-apolipoprotein A-I • ProteinChip

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