Patent ID: 6805799
Filing Date: 2004-10-19
Classification: B01D

Abstract:
A process for continuously separating and focusing components from a mixture, said process comprising:(1) forming a F section for chromatographic focusing by arranging f zones in series where f is an integer and is larger than four, said zones being sequentially numbered from zone F(f) to zone F(1) wherein zone F(f) is arbitrarily defined as the upstream end and zone F(1) is arbitrarily defined as the downstream end, each zone having an incoming joint fluidly connecting with an inlet tube and an outgoing joint fluidly communicating with an outlet tube, a column of chromatography being detachably installed in-between the incoming joint and the outgoing joint of each zone, said columns having a stationary phase chromatographic stationary phases that include ion exchange, affinity, chelating, chiral, ligand exchange, size exclusion, monolithic, polymeric, normal phases, and having a stationary phase, each inlet tube and each outlet tube having an additional branched opening and the branched opening of the outlet tube of one zone being connected with the branched opening of the inlet tube of the adjacent downstream zone to form a fluid passage between the two zones, the branched opening of the inlet tube of zone F(f) and the branched opening of the outlet tube of zone F(1) being sealed or eliminated during tube manufacturing to allow a fluid entering the inlet tube of zone F(f) and exiting the outlet tube of zone F(1); (2) selecting at least four inlet tubes and at least two outlet tubes in the F section for fluid delivery and fluid withdrawal with the requirement that at least one fluid passage has both the inlet tube and the outlet tube selected, blocking the unselected outlet tubes and the unselected inlet tubes, determining fluid compositions and flow rates for each of the selected outlet tubes and the selected inlet tubes to generate a desired desorption gradient, hydrophobic gradient, pH gradient, chelating gradient, hydrogen bonding gradient, affinity gradient, chiral gradient and other gradients for said unselected inlet tubes and said unselected outlet tubes being optionally built without the branched openings so that they can replace the branched openings to form the fluid passages; (3) performing chromatographic focusing in the F section by: (a) continuously and simultaneously delivering and withdrawing the following fluids of defined compositions at defined flow rates through the selected inlet tubes and the selected outlet tubes with pumps, including: delivering an eluent of high desorption strength into the inlet tube of zone F(f); delivering a sample solution into at least one selected inlet tube; delivering focusing solutions of lower desorption strengths into the remaining selected inlet tubes, a focusing solution being delivered through the inlet tube of one zone and being mixed with the eluent from its adjacent upstream zone to form a different eluent of lower desorption strength that flows into said zone, a desorption gradient being thus generated in the F section with the eluent of zone F(f) having the highest desorption strength and the eluent of zone F(1) having the lowest desorption strength, there being at least four levels of desorption strengths among the zones of the F section; collecting the flow-out fluid from the outlet tube of zone F(1), withdrawing fluids from the remaining selected outlet tubes at a flow rate smaller than 50% of the flow rate for the eluent; (b) periodically and simultaneously relocating the column in zone F(f) to zone F(1) and relocating the rest columns one zone upstream for continuous chromatographic focusing, components of a sample being introduced in a zone, absorbed on the column in the zone, brought zone by zone upstream by column relocations, selectively desorbed from different zones due to the differences in desorption strengths, and separated from each other, a component desorbed from a zone being eluted back to the adjacent downstream zone where said component is reabsorbed on the column of said adjacent downstream zone that contains newly introduced sample, said component being again brought to said zone by the next column relocation and being thus retained and accumulated between said zone and said downstream zone by continuous sample delivery and repeatedly column relocation.