University of Illinois Urbana-Champaign

The Effect of Immunoaffinity Column Chromatography Elution Buffers on HDL Apolipoprotein Detection

Luna, Viridiana

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https://hdl.handle.net/2142/95802

Description

Title
The Effect of Immunoaffinity Column Chromatography Elution Buffers on HDL Apolipoprotein Detection
Author(s)
Luna, Viridiana
Contributor(s)
Andrade, Flavia
Issue Date
2016
Keyword(s)
Molecular and Cellular Biology
Abstract
High-density lipoproteins (HDL) are complex particles composed of lipids and apoliproteins. The major function of HDL is to transport cholesterol from peripheral tissues to the liver. HDL apolipoproteins include apoAI, a major structural apolipoprotein; apoCIII, a protein that inhibits binding of apoE and apoB-100 to hepatic receptors; and apoE, a ligand for the low-density lipoprotein (LDL) receptor, apoE receptor, and LDL receptor related protein. Immunoaffinity column chromatography methods are frequently used to isolate HDL subspecies. Therefore, it is important to understand how elution buffers may be affecting protein detection and potentially denaturing proteins. In order to optimize immunoaffinity column chromatography elution conditions, we tested the effects of two elution buffers, acetic acid and sodium thiocyanate (NaSCN), on apoA1, apoCIII, and apoE. We tested the elution buffers for: number of elutions required to elute protein, protein recovery, and reduction of protein detection after treatment with elution buffer. Plasma samples containing known concentrations of apoliproteins listed were used. This allowed us to compare expected concentrations and measured concentrations after treatment. The sandwich enzyme-linked immunosorbent assay (ELISA) method was used to quantify protein concentrations. The results of these experiments will allow us to maximize protein recovery and account for loss of protein detection due to treatment with elution buffers.
Publisher
Office of Minority Student Affairs
Type of Resource
text
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http://hdl.handle.net/2142/95802
Copyright and License Information
Copyright 2016 Viridiana Luna

Owning Collections

TRiO - Vol. 2, no.1 2016 PRIMARY
The TRiO McNair journal is a culmination of research conducted by student scholars and their facutly representatives through the Ronald E. McNair Scholars Program.