A reciprocating free-flow isoelectric focusing electrophoresis device and method

Abstract

The invention discloses a reciprocating type free flow isoelectric focusing electrophoresis device and a reciprocating type free flow isoelectric focusing electrophoresis method. The reciprocating type free flow isoelectric focusing electrophoresis device is mainly composed of a separation chamber, a liquid storage pipe, a gas liquid buffering chamber, collection pipes, recycling pipes which are located below the collection pipes and correspond to the collection pipes one by one, and a medium driving pump, wherein a separation chamber liquid outlet of the separation chamber is connected with the collection pipes and the recycling pipes respectively by switches; the liquid storage pipe is connected with a gas liquid buffering chamber liquid outlet; the gas liquid buffering chamber liquid outlet is connected with the separation chamber liquid outlet; and the medium driving pump drives media to flow in or out of the gas liquid buffering chamber from the gas liquid buffering chamber liquid inlet. According to the device and the method, one single-path driving pump is used for replacing a multi-channel driving pump; the rapid reciprocating of CA (Carrier Ampholytes) buffering liquid and a sample is used for replacing unidirectional circulation in the prior art by the separation chamber, so that the same effect of an existing RFFIEF technology is achieved; and the technology, equipment and method of the whole ReFFIEF technology are greatly simplified and the cost is reduced.

Description

technical field [0001] The invention relates to a preparative electrophoresis device and method in the field of biochemical technology, in particular to a reciprocating free-flow isoelectric focusing electrophoresis device and method. It is mainly used in free-flow isoelectric focusing electrophoresis separation of proteins, polypeptides, cells, organelles, viruses and other amphipathic substances and particles. Background technique [0002] Freeflow electrophoresis (FFE) is a pure liquid-phase electrophoresis technique with both preparation and analysis functions. It has the advantages of mild separation conditions, high recovery rate, and continuous separation and purification. Since Hanning invented free flow electrophoresis in 1961 (Hanning, K., Fresenius J. Anal. Chem. 1961, 181, 244-254), FFE has developed a variety of electrophoretic preparation techniques, such as free flow zone electrophoresis (Freeflow zone electrophoresis, FFZE), Free flow isoelectric focusing el...

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Problems solved by technology

[0006] However, the above-mentioned RFFIEF technology has the following problems: first, the device is complex and expensive, requiring expensive multi-channel drive pumps; second, the number of hoses that can be driven by the multi-channel drive pumps is limited, which limits the design of the device; third, the operation is complicated , the introduction and recovery of the solution are troublesome, and the gas discharge from the separation chamber needs to be repeatedly adjusted to the rotation direction of the multi-channel drive pump to achieve

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