Figure 1.

Schematic representation of the platform for the simultaneous depletion of albumin and Igs, and the enrichment of fucosylated glycoproteins, and the subsequent RPC fractionation of the captured proteins. When the switching valve (SV) A, B, C and D were in 2, 3, 5 and 7 positions, respectively, the 3-fold diluted serum was injected onto the depletion and the lectin columns, followed by washing with the binding mobile phase (BMP) using pump A. Then, the eluting mobile phase for the depletion columns was passed by changing the SV-A position to 1, thus by-passing the lectin and the RPC columns. The depletion columns were re-equilibrated again with the BMP, after which the SV-A was changed back to position 2. Then, the LTA column was eluted using pump B, while the 3-way valve was in position 9, SV-B in position 3 and SV-C in position 6, thus by-passing the AAL column and passing through the RPC column. This was followed by washing, eluting and re-equilibrating the RPC column using the mobile phase from pump B, while the 3-way valve is in position 10. Then, the AAL column was eluted by changing the 3-way valve position back to 9, SV-B position to 4, SV-C position to 5 and SV-D position to 8. This was again followed by washing, eluting and re-equilibrating the RPC column by keeping the 3-way-valve in position 10.