This document describes a new technology for producing polymer monolithic liquid chromatography columns with very low internal diameters down to 0.02mm. These narrow columns allow for more sensitive separation and identification of peptides using LC-MS/MS. The technology involves a specific preparation process for the monomers, porogens and initiator that allows creation of monolithic columns that are difficult to produce at these small sizes. Initial testing showed the columns can successfully separate peptides from cerebrospinal fluid tryptic digests. Potential applications include both clinical diagnostics and research due to the increased sensitivity and reproducibility of preparation compared to existing options.

1. POLYMER MONOLITHIC COLUMN AND
PREPARATION TECHNOLOGY
Keywords: Liquid Chromatography, monolithic column, peptide separation, bioinformatics, lab-on-a-chip,
diagnostic array
Background
The Achilles heel of bottom-up proteomics is the overwhelming number of peptides that need
to be separated and identified using liquid chromatography coupled with tandem mass
spectrometry (LC-MS/MS). Sensitivity can be improved by using chromatography columns
with low internal diameter (ID). Because of the difficulty in filling a very narrow capillary tube
with particles, commercially available packed columns are technologically limited to 0.075 mm
ID.
Description
In theory, there is no such limitation to preparation of monolithic columns. Still, there are few
monolithic products available, and typically the ID’s of these are 0.1 mm and higher. The
preparation of very narrow ID monolithic columns is difficult. The product is a preparation
technology (the ingredients - monomers, porogens and initiator - and the preparation setup) of
polymer monolithic liquid chromatography column with very low internal diameter (0.03 mm)
for sensitive nanoscale peptide separations.
Figure BPI chromatogram of peptides from CSF tryptic digest using polymeric monolith and trap-column
Applications and benefits
- Very low internal diameter (ID is between 0,05mm and 0,02mm)
- Easy to reproduce – lower preparation costs, time and difficulty
- Opportunities for both clinical diagnostic and R&D
Status
Verified and tested prototype
IPR
Patent pending
We are seeking licensing partners.