System and method for the separation of analytes
View Patent ↗A separation module operates to fractionate or separate an analyte into fractions according to pI, i.e., pI bands, utilizing capillary isoelectric focusing (“CIEF”) within a first microchannel. The fractions are stacked to form plugs, the number of which is determined by a number of parallel second microchannels integrally connected to the first microchannel, into which the fractions are directed according to the buffer characteristics found in each of the individual microchannels. Within the microchannels the plugs are separated into proteins according to a different chemical property, i.e., “m/z,” utilizing capillary electrophoresis (“CE”).
1. A system for separating a sample comprising:
injection adapters that introduce a sample into a first microchannel formed in a module;
a first microchannel of the module that separates the sample into a plurality of sub-samples based on varying isoelectric points of the plurality of sub-samples, the first microchannel having an isoelectric focusing buffer with a first ionic strength formed therein;
at least one second microchannel that separates a first of the plurality of sub-samples into a plurality of protein components according to electrophoresis, the at least one second microchannel having an electrophoresis buffer with a second ionic strength therein, wherein the second ionic strength of the electrophoresis buffer is different from the first ionic strength of the isoelectric focusing buffer;
electrodes that apply opposite charges to either end of the first microchannel to facilitate separating the sample into the plurality of sub-samples;
a translator that aligns a first detector with the first microchannel to detect the separation of the sample into the plurality of sub-samples based on varying isoelectric points of the plurality of sub-samples within the first microchannel, and aligns a second detector with one microchannel of the at least one second microchannel.
2. The system according to claim 1 , wherein the first detector comprises UV-Visible (“UV-Vis”) and photodiode (“PDA”) arrays.
3. The system according to claim 2 , wherein the UV-Vis and PDA arrays continuously monitor the separation of the sample into the plurality of sub-samples.
4. The system according to claim 1 , wherein each of the plurality of sub-samples contains at least one known component and at least one unknown component.
5. The system according to claim 4 , wherein the at least one known component is an internal standard and the at least one unknown component is a protein component.
6. The system according to claim 5 , wherein the protein component is at least one of an amino acid, a peptide, a polypeptide or a protein.
7. The system according to claim 1 , wherein the second detector is a mass spectrometer that produces a spectrum indicative of each of the plurality of protein components.