Introduction to Tangential Flow Filtration

タンジェンシャルフロー・フィルトレーション(TFF)は、生体分子を迅速かつ効率よく分離および精製する方法のひとつです。免疫学、タンパク質化学、分子生物学、生物化学、微生物など幅広い生物学分野に利用できます。TFF は、10mL から数千L までの幅広いボリュームの試料液の濃縮および脱塩に使用されます。生体分子のサイズを問わず、分画または細胞懸濁液からのセルハーベスト、醗酵ブロスや細胞溶解液の清澄化などの用途に最適です。This report describes the basic principles that govern TFF and the use of TFF capsules and cassettes in laboratory and process development applications.

タンジェンシャルフロー・フィルトレーションの概要

Membrane filtration is a separation technique widely used in the life science laboratory.Depending on membrane porosity, it can be classified as a microfiltration or ultrafiltration process.Microfiltration membranes, with pore sizes typically between 0.1 µm and 10 µm, are generally used for clarification, sterilization, and removal of microparticulates or for cell harvesting.Ultrafiltration membranes,
 







with much smaller pore sizes between 0.001 and 0.1 µm, are used for concentrating and desalting dissolved molecules (proteins, peptides, nucleic acids, carbohydrates, and other biomolecules), exchanging buffers, and gross fractionation.Ultrafiltration membranes are typically classified by molecular weight cutoff (MWCO) rather than pore size.

There are two main membrane filtration modes which can use either microfiltration or ultrafiltration membranes:1) Direct Flow Filtration (DFF), also known as ”dead-end” filtration, applies the feed stream perpendicular to the membrane face and attempts to pass 100% of the fluid through the membrane, and 2) Tangential Flow Filtration (TFF), also known as crossflow filtration, where the feed stream passes parallel to the membrane face as one portion passes through the membrane (permeate) while the remainder (retentate) is recirculated back to the feed reservoir.

TFFの理論を理解するための良い例えは、選別スクリーンを使用して小石から砂の分離を試みる場合などが挙げられます。スクリーンの穴はメンブレンの孔であり、砂と小石は分離される分子です。In DFF, the sand and pebble mixture is forced toward the holes in the screen.The smaller sand grains fall through the pores in the screen, but the larger pebbles form a layer on the surface of the screen.This prevents sand grains at the top of the mixture from moving to and through the holes (Figure 1A).DFFでは、圧力が高まることで、分離を増やすことなく単純に混合物が圧縮されます。In contrast, operating in a TFF mode prevents the formation of a restrictive layer by re-circulating the mixture.The process acts like a shaking sifter to remove the pebbles that block the holes in the screen, allowing the sand grains at the top of the mixture to fall toward and through the holes in the screen (Figure 1B).

図 1

選別スクリーンを使用した砂と小石の分離

TFF:選別スクリーンを使用した砂と小石の分離
(A) Applying direct pressure to the mixture allows the sand grains at the bottom to fall through.A layer of pebbles builds up at the screen surface preventing sand grains at the top from moving to and through the screen.
(B)Shaking the screen breaks up the aggregated pebble layer at the bottom of the mixture and allows for complete fractionation.The crossflow dynamic of the feed stream in tangential flow filtration serves the same purpose as shaking in this example.

In solution, the same effect is encountered for DFF (Figure 2) and for TFF (Figure 3).The flow of sample solution across the membrane surface sweeps away aggregating molecules that form a membrane-clogging gel (gel polarization), allowing molecules smaller than the membrane pores to move toward and through the membrane.Thus, TFF can be faster and more efficient than DFF for size separation. 

図2

Direct Flow Filtration Process

TFF:Direct Flow Filtration Process
(A) 供給がメンブレンに向かいます。Molecules larger than the pores accumulate at the membrane surface to form a gel, which fouls the surface, blocking the flow of liquid through the membrane.
(B)As the volume filtered increases, fouling increases and the flux rate decreases rapidly.

図3

Tangential Flow Filtration Process

Tangential Flow Filtration Process
(A) サンプル液が、供給チャンネルからメンブレンの表面に沿って (接して)、メンブレンからも流れています。The crossflow prevents build up of molecules at the surface that can cause fouling.
(B)The TFF process prevents the rapid decline in flux rate seen in direct flow filtration allowing a greater volume to be processed per unit area of membrane surface.