DIAFILTRATON洗滤.doc

Scientific Technical Report PN 33289 Diafiltration: A Fast, Efficient Method for Desalting, or Buffer Exchange of Biological Samples By Larry Schwartz, Senior Technical Manager, Pall Life Sciences  Diafiltration is a technique that uses ultrafiltration membranes to completely remove, replace, or lower the concentration of salts or solvents from solutions containing proteins, peptides, nucleic acids, and other biomolecules. The process selectively utilizes permeable (porous) membrane filters to separate the components of solutions and suspensions based on their molecular size. An ultrafiltration membrane retains molecules that are larger than the pores of the membrane while smaller molecules such as salts, solvents and water, which are 100% permeable, freely pass through the membrane. This article will cover the concepts of protein concentration and diafiltration. It will compare different ways of performing diafiltration and their impact on process time, volume, product stability, and recovery. CONCENTRATION The solution retained by the membrane is known as the concentrate or retentate. The solution that passes through the membrane is known as the filtrate or permeate. A membrane for concentration is selected based on its rejection characteristics for the sample to be concentrated. As a general rule, the molecular weight cut-off (MWCO) of the membrane should be 1/3rd to 1/6th the molecular weight of the molecule to be retained (3-6X Rule). This is to assure complete retention. The closer the MWCO is to that of the sample, the greater the risk for some small product loss during concentration. The risk increases if diafiltration will also be used since the relative loss depends on the total volume of filtrate that will be generated. Membrane flux rate (filtrate flow rate per unit area of membrane) is related to pore size. The smaller the pores, the lower the flux rate for the same applied pressure. Therefore, when selecting a membrane for concentr