Phospholipid nano-disk chromatography medium and preparation method and application thereof

Abstract

The invention relates to a phospholipid nano-disk chromatography medium and a preparation method and application thereof. The phospholipid nano-disk chromatography medium comprises a charged phospholipid nano disk and an affinity matrix; the charged phospholipid nano disk comprises phospholipid and a film scaffold protein with an affinity label; the charged phospholipid nano disk is connected withan affinity matrix through the affinity label. The phospholipid nano-disk chromatography medium has both binding capacity and protection capacity to film proteins, so that the actual system containing the film proteins can be purified and reconstructed on a chromatography column in one step; and the phospholipid nano-disk chromatography medium has reversibility and regeneration performance; the phospholipid nano-disk chromatography medium solves the problems of serious protein activity loss, long and complicated steps and time-consuming and labor-consuming operation in the traditional film protein research technology, is simple and efficient to operate, improves the film protein activity recovery rate by more than 20 times, and shortens the treatment time by more than 95%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a chromatographic medium for separating and purifying membrane proteins, in particular to a phospholipid nanodisc chromatographic medium and its preparation method and application. Background technique [0002] Membrane proteins, as the main bearers of biomembrane functions, not only provide a stable internal environment for the life activities of individual cells, but also mediate the connection between cells and cells, and between cells and substrates, thus playing an important role in the signal transduction and physiological transport of living organisms. , Enzyme catalysis and structural connection play a rich and important function. Therefore, membrane proteins have always been the focus of research in the fields of biology, medicine and materials. However, membrane proteins are also recognized as research difficulties. The vast majority of membrane proteins are e...

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Problems solved by technology

[0004] In the prior art, the traditional research method of first purifying membrane proteins and then remodeling them has the following problems: 1. No matter what kind of remodeling form it is, it only has a protective function on membrane proteins, but has no binding ability. The purification and reconstruction of membrane proteins is a fragmented process, which makes the whole operation process cumbersome, time-consuming and laborious; 2. The activity loss is large. Taking plasma membrane proteins as an example, after a series of purification steps, the protein activity is only 30 %, the reconstitution process can only increase its activity to 60% at most, that is to say, the membrane protein activity will still lose nearly half of the membrane protein activity by adopting this step-by-step treatment method of purification and reconstitution; 3. The research area is narrow and the reconstitution The structured membrane protein is bound to the surface of the matrix and cannot be peeled off. The follow-up can only be studied by a specific research technique for this type of matrix, and the amount of information obtained is small.

[0005] At present, there is no chromatographic system that can realize membrane protein purification and reconstitution in one step in the prior art. Based on this, it is urgent to develop a new membrane protein research strategy to achieve efficient purification and activity maintenance of membrane proteins at the same time. Target

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