Patent ID: 9566537
Date: 2017-02-14
CPC Classifications: B01D,C07K,G01N

Claim:
1. A multidimensional liquid chromatography separation system for protein separation, the multidimensional liquid chromatography separation system comprising a detection device, a mobile phase tank, a first infusion device, a second infusion device, a first injector device, a second injector device, a separation device, a collect-reserve device comprising at least one sample loop, at least two draining devices, and a flow path switching device, wherein: the mobile phase tank is used for the storage of mobile phase for the multidimensional liquid chromatography separation; the first and second infusion devices are employed independently to take out mobile phase which is suitable for the liquid chromatography separation from the mobile phase tank and then, the mobile phase is transported to the multidimensional liquid chromatography separation system; and the first infusion device and the second infusion device measure and adjust the flow rate of the mobile phase independently; in the first dimensional separation, the first infusion device is used to transport mobile phase suitable for the first dimensional separation to the first injector device; in the second dimension or a higher dimensional separation, the first infusion device is used to transport mobile phase suitable for the second dimensional or the higher dimensional separation to the collect-reserve device and push the intermediate fraction stored in the collect-reserve device into the second injector device; at the same time, the second infusion device takes out mobile phase suitable for the second dimensional or higher dimensional separation from the mobile phase tank and then, transport it to the second sample injection device; the first injection device introduces original samples from outside of the system and pushes the original samples into the separation device together with mobile phase from the first infusion device; the second injector device includes a sample mixer; said sample mixer is used to mix the previous fraction from at least one sample loop and mobile phase from the collect-reserve device and mobile phase from the second infusion device, so that a sample mixture is obtained, and then, the sample mixture is pushed into a next dimensional separation device; the separation device includes a chromatographic column switching unit and a total number, “n” chromatographic columns, or a chromatographic column with a total number of separation modes “m”, and liquid in the separation device is separated into different fractions through the chromatographic column, in which the chromatographic column selector helps to put the liquid in the separation device selectively into one of the chromatographic column of the “n” chromatographic columns; “n” is a nonnegative integer and “m” is the number of different separation modes used in the multidimensional liquid chromatography separation system, and “n” and “m” need to satisfy:  wherein “i” is a positive integer; “q” stands for the number of chromatographic columns having the number “i” of separation modes; the collect-reserve device includes the number “p” of the sample loops switching units and “p” is greater than or equal to 1; the sample loops switching unit is used to control the direction of liquid movement in the collection and collect-reserve device to selectively pass through at least one of the collect-reserve devices and then, the collect-reserve device is employed to collect the intermediate fraction that has been separated and required to do subsequent separation by other separation mode, and the second fraction containing the target protein is also stored in at least one of the collect-reserve devices; said at least two draining devices push the liquid in the flow path out of the system; the flow path switching device is made up from valves connected to the device and pipes above by switching the valves of the flow path switching device, it provides not only the flow path for conventional liquid chromatography separation but also the flow path for multidimensional liquid chromatography separation; and the first infusion device and the second infusion device adjust and measure the flow rate of mobile phase for the second or higher dimensional separation and adjust an eluent concentration of the sample mixture in the second or higher dimensional separation to be lower than a critical migration eluent concentration C