Shakya Akhilesh Kumar, Srivastava Akshay, Kumar Ashok
Department of Chemical Engineering, Texas Tech University, Lubbock, Texas.
Network of Excellence for Functional Biomaterials, National University of Ireland, Galway, Republic of Ireland.
Curr Protoc Nucleic Acid Chem. 2015 Dec 1;63:10.16.1-10.16.10. doi: 10.1002/0471142700.nc1016s63.
Three-dimensional monolithic columns are preferred stationary phase in column chromatography. Conventional columns based on silica or particles are efficient in bioseparation though associated with limitations of nonspecific interaction and uneven porosity that causes high mass transfer resistance for the movement of big molecules. Cryogels as a monolith column have shown promising application in bioseparation. Cryogels column can be synthesized in the form of a monolith at sub-zero temperature through gelation of pre-synthesized polymers or polymerization of monomers. Cryogels are macroporous and mechanically stable materials. They have open interconnected micron-sized pores with a wide range of porosity (10-200 μm). Current protocol demonstrated the ability of poly(hydroxymethyl methacrylate)-co-vinylphenyl boronic acid p(HEMA-co-VPBA) cryogel matrix for selective separation of RNA from the bacterial crude extract.
三维整体柱是柱色谱中首选的固定相。基于硅胶或颗粒的传统柱在生物分离中效率较高，尽管存在非特异性相互作用和孔隙率不均匀的局限性，这会导致大分子移动时传质阻力较高。作为整体柱的冷冻凝胶在生物分离中已显示出有前景的应用。冷冻凝胶柱可以通过预合成聚合物的凝胶化或单体的聚合在零下温度以整体形式合成。冷冻凝胶是大孔且机械稳定的材料。它们具有开放的相互连接的微米级孔隙，孔隙率范围很广（10 - 200μm）。当前方案证明了聚（甲基丙烯酸羟甲酯）- 共 - 乙烯基苯基硼酸p（HEMA - co - VPBA）冷冻凝胶基质从细菌粗提物中选择性分离RNA的能力。
