Proteomics reactor, protein chromatographic separation platform and use thereof

A chromatographic separation device for preparing a sample for proteomics analysis, comprising:

• (a) a sample preparation device which comprises:
• (1) a pipette tip of about 200 uL;
• (2) a solid-phase extraction membrane with a length of about 3 mm filled at the bottom end of the pipette tip;
• and (3) a layer of strong cation exchange resin or strong anion exchange resin located above the solid-phase extraction membrane, wherein the sample preparation device does not comprise a second solid-phase extraction membrane located above the layer of strong cation exchange resin or strong anion exchange resin;
• (b) a lysis buffer for lysing a biological sample into a plurality of proteins, comprising HEPES, NaCl, guanidine hydrochloride, n-dodecyl β-D-maltoside (DDM), and a protease inhibitor;
• (c) a digesting reagent for digesting the plurality of proteins retained in the layer of strong cation exchange resin or strong anion exchange resin into a plurality of peptides, comprising trypsin;
• (d) a transfer buffer for transferring the plurality of peptides in the layer of strong cation exchange resin or strong anion exchange resin into the solid-phase extraction membrane, comprising sodium chloride, ammonium formate or ammonium bicarbonate;
• and (e) an elution buffer for eluting the plurality of peptides from the solid-phase extraction membrane, comprising (1) ammonium formate or acetic acid, and (2) acetonitrile.