Process for industrially separating biopolymers

Claim 1

A process for controlling the industrial separation of a biopolymer by column chromatography, in which a large amount of a biopolymer-containing mixture sample is charged into the upper stream side of a column filled with a filler having ion-exchange capability; a mobile phase liquid the concentration and/or pH of which continuously vary/varies with the passage of time is prepared by continuously mixing at least 2 precursor liquids for the mobile phase while controlling the flow rate of each precursor liquid over a wide variable range of flow rate; the mobile phase liquid thus obtained is pumped through the column to develop the components of the mixture in the column; and the fractions eluted from the lower stream side of the column are collected to separate the components of the mixture from each other, said process comprising sequentially conducting the following steps: (i) a step of pumping a buffer solution through a column in which a filler having ion-exchange capability has been charged to bufferize the column until it is confirmed that the observed value of the eluate from the column has been maintained within a predetermined range for a predetermined time after passage of a specific period of time from the pumping of the liquid; (ii) a step of injecting a biopolymer-containing mixture sample into the column and finishing after passage of a specific period of time from the injection; (iii) a step of pumping a washing liquid into the column until it is confirmed that the observed value of the eluate from the column measured by a detector has been maintained within a predetermined range for a predetermined time after passage of a specific period of time from the pumping of the liquid; (iv) a step of preparing a mobile phase liquid the concentration and/or pH of which are/is continuously varied by controlling the ratios of pumping flow rate of respective liquids for mobile phase according to a predetermined program, pumping the mobile phase liquid into the column to develop the components of the sample within the column and collecting the eluate from the lower stream side of the column in fractions so that components of the sample are separated from each other, in which fractionation is started after passage of a specific period of time from the start of the process or when the observed value exceeds the predetermined value; (v) a step of pumping a washing-out liquid into the column to wash out unnecessary components from the column until it is confirmed that the observed value of the eluate from the column measured by a detector has been maintained within a predetermined range for a predetermined time after passage of a specific period of time from the pumping of the liquid; (vi) a step of pumping regenerating liquids having different concentrations sequentially into the column until it is confirmed that the observed value of the eluate from the column measured by a detector has been maintained within a predetermined range for a predetermined time after passage of a specific period of time from the pumping of the liquid; and (vii) a step of pumping a cleaning liquid into the column for a predetermined time.