Preparation of lipidosome containing MAP30 protein and entrapping method

Abstract

The invention provides preparation of lipidosome containing MAP30 protein and an entrapping method. A thin film hydration method is used for preparing. The preparation specifically comprises the following steps of dissolving 10mg of DPPC (dipalmitoyl phosphatidyl choline), 1mg of cholesterol, 1mg of n-(carbonyl-methoxypolyethyleneglycol 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (SDPE-mPEG2000) into chloroform, performing pressure relief and rotary evaporation to form a uniform thin film, thoroughly volatizing the remained chloroform overnight under the vacuum condition, adding 1ml of 2mg ml<-1> PBS (poly(butylene succinate)) solution containing protein, performing ultrasonic treatment for 20min in an ice bath, and enabling the lipidosome film to fall down; oscillating for 5min on an oscillator, fully hydrolyzing, and forming turbid liquid; transferring the turbid liquid into an EP (electrochemical polishing) tube, using a probe to perform ultrasonic treatment for 30min at the power equal to 15% of 135W ultrasonic power, so as to obtain a transparent and uniform blue lipidosome suspension. The preparation has the advantages that the technology is simple, the conditions are mild, the repeatability is high, the structure of the lipidosome is stable, the protein adsorbing capacity is large, the particle sizes of the lipidosome are more uniform, and the encapsulating rate is 78%.

Description

technical field [0001] The invention relates to a method for preparing and encapsulating liposomes containing MAP30 protein. Background technique [0002] MAP30, the type I ribosome inactivating protein of bitter melon, is a protein against acquired immunodeficiency syndrome virus with a relative molecular mass of 30,000 in bitter melon. It belongs to the single chain type I ribosome inactivating protein (ribosome inhibiting protein, RIP) family. It has antiviral, antitumor, antibacterial and other biological activities, and has become a hot spot in the research of anticancer drugs in recent years. Type I ribosome inactivating protein is a single-chain protein that cannot enter normal cells. MAP30 is a highly toxic protein that can produce cytotoxicity to cells including normal cells before it enters cells. [0003] The above-mentioned problems should be considered and solved in the preparation and encapsulation process of liposomes. Contents of the invention [0004] Th...

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Problems solved by technology

[0004] The object of the present invention is to provide a method for preparing and encapsulating liposomes containing MAP30 protein to solve the problem that the type I ribosome inactivating protein in the prior art is a single-chain protein that cannot enter normal cells, and MAP30 is a relatively toxic liposome. Large protein, before it enters the cell, it will cause cytotoxicity to the cell, including normal cells

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