| 摘要 |
A multidimensional liquid chromatography separation system has a mobile-phase storage tank, a first liquid transfer device, a second liquid transfer device, a first sample introduction device, a second sample introduction device, a separation device, a collection, storage device, at least two drainage devices and a flow path switching device. A separation method for protein separation using the multidimensional liquid chromatography separation system has the steps of: 1) preparation in advance, 2) the first dimensional separation, 3) collection and storage of the intermediate fraction, 4) the second dimensional or multidimensional separation and repeating the steps 3) and 4). |
| 主权项 |
1. A multidimensional liquid chromatography separation system for protein separation, comprising a detection device, wherein the multidimensional liquid chromatography separation system comprises a mobile phase tank, a first infusion device, a second infusion device, a first injector device, a second injector device, separation device, collect-reserve device comprising at least one sample loop, at least two draining devices and flow path switching device, in which:
1) the mobile phase tank is used for the storage of mobile phase for the multidimensional liquid chromatography separation; 2) 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 they measure and adjust their flow amount 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 multidimensional separation, the first infusion device is used to transport mobile phase suitable for the second dimensional or multidimensional 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 more multidimensional separation from the mobile phase tanks and then, transport it to the second sample injection device; 3) introducing original samples from outside the system and pushing them into the separation device together with mobile phase from the first infusion device; the second injector device includes 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; 4) the separation device includes 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:
m≧n (1)m−q×(i−1) (2)wherein “i” is a positive integer; “qi” stands for the number of chromatographic columns having the number “i” of separation modes;
5) 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; 6) said at least two draining devices push the liquid in the flow path out of the system; and 7) 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 further characterized in that the first infusion device and the second infusion device can adjust and measure the flow amount of mobile phase they transport for the second or more dimensional separation so as to adjust the eluent concentration of the injection mixture using in the second or multidimensional separation, when sampling, the eluent concentration of the injected mixture is lower than the critical migration eluent concentration on which all the target protein in the target intermediate fractions must be retained in the second or more dimensional separation. |
