Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins  

Miriam S. Goyder¹, Keith R. Willison², David R. Klug¹, Andrew J. deMello^3,* & Oscar Ces^1,*  

¹The Single Cell Proteomics Group, Institute of Chemical Biology, Department of Chemistry, Imperial College London, South Kensington, London, UK SW72AZ, ²Institute of Cancer Research, Chester Beatty Laboratories, Section of Cell and Molecular Biology, London, UK SW3 6JB, ³Institute of Chemical and Biochemical Engineering, ETH Z  

We present a top down separation platform for yeast ribosomal
proteins using affinity chromatography and capillary electrophoresis
which is designed to allow deposition of proteins onto
a substrate. FLAG tagged ribosomes were affinity purified, and
rRNA acid precipitation was performed on the ribosomes followed
by capillary electrophoresis to separate the ribosomal
proteins. Over 26 peaks were detected with excellent reproducibility
(＜0.5% RSD migration time). This is the first reported
separation of eukaryotic ribosomal proteins using capillary
electrophoresis. The two stages in this workflow, affinity chromatography
and capillary electrophoresis, share the advantages
that they are fast, flexible and have small sample requirements
in comparison to more commonly used techniques. This method
is a remarkably quick route from cell to separation that has
the potential to be coupled to high throughput readout platforms
for studies of the ribosomal proteome.  

Affinity chromatography, Capillary electrophoresis, EVV 2DIR, Proteomics, Ribosomal proteins