切向流过滤:更简便、更快捷、更高效

当需要高效、经济地去除杂质且涉及大量样品时, 切向流过滤 (TFF) 可能是最佳选择。TFF 采用 过滤器 分离或隔离特定粒径的颗粒。不同于传统(或死端)过滤,切向流过滤允许流体循环穿过过滤器以产生压差,而不是强制流体通过。

这种 “切向”过滤 方式意味着样品平行流过过滤器,并通过储液器进行多次循环。小于孔径的颗粒可通过滤膜,形成滤液。 TFF也称为 错流过滤,包括微滤、纳滤和超滤。

切向流过滤的优势

使用 TFF,可避免较大颗粒在过滤器中堆积或结块,提高了过滤器的可用性。其他 优势 包括:

使用简便:

设置完成(包括泵、压力表、泵管和配件,如下图所示)且将样品放入储液罐后,系统便可运行

高效率运作:

TFF 可以连续运行(不用分批进行)并快速分离和提纯生物分子

节省时间:

样品浓缩和渗滤可在同一系统上完成

可处理大样本或小样本:

切向流过滤产品可用于处理 10 mL 到数千升的样品 Masterflex Peristaltic Pump System for TFF

TFF 应用

一位研究人员想研究寄生在牛肠道上的病毒。他创建了一个系统以进行研究,这个系统组成包括 Masterflex® L/S® 蠕动泵、带有硅胶管的泵头、压力表以及用于仪表安装和过滤器连接的接头。他通过蠕动泵和压力表将样品从储液罐中输送到 切向流过滤器中。他在不同的流程中使用了两个过滤器:一个用于去除细菌的 0.2 µm 过滤器和一个用于浓缩病毒的 <100 kD 过滤器。

结果显示,与其他过滤控制水平较低的系统相比,该系统采用切向流过滤器捕获的病毒浓度更高。

在葡萄酒行业,切向流过滤并不是一项新技术,用以去除细胞碎片,生产更清洁的葡萄酒,且节省了装瓶前的终滤成本。葡萄酒本身的质量也有所提高。TFF 在食品加工领域的应用长达数十年,当用于药物加工和制造时,可以连续提纯蛋白质。

何时采用切向流过滤

TFF 适用于较大表面积,并且可过滤较大体积的物质而不会发生堵塞。TFF 是大规模实验室分离的理想选择,例如细胞收获或从培养上清液(沉淀物或沉淀物上方的透明液体)中分离蛋白质。具体应用包括:

  • 回收和提纯抗体、质粒 DNA 或重组蛋白
  • 收获细胞
  • 制备色谱样品
  • 浓缩和脱盐蛋白质和肽或核酸 (DNA)
  • 分馏稀释的蛋白质混合物
  • 去除水、缓冲液和培养基溶液中的内毒素
  • 净化发酵液

推荐的切向流过滤设备

  • Masterflex® 蠕动泵(例如 Masterflex® L/S® 数字模块化分配驱动器或 Masterflex® I/P® 数字过程驱动器)和泵头,满足所有需求且能保持样品完整性。采用双通道泵头,可同时运行两个过滤器
  • 压力表
  • Masterflex® 泵管(如 Masterflex® 硅胶管)
  • 用于仪表安装和过滤器连接的接头
  • 切向流过滤器

In fields such as biochemistry, immunology and microbiology, the separation and purification of biological material is paramount to the forward march of innovation.

As such, lab experts have developed tangential flow filtration, which succeeds at:

  • Removing cells from cell culture broths
  • Diafiltration
  • Fractionation
  • Concentration

What is Tangential Flow Filtration?

Tangential flow filtration (TFF) is an efficient, cost-effective way of removing impurities from large volume samples. It involves using a filter to separate or isolate contaminant particles. Unlike dead-end filtration, in TFF fluid is circulated across, rather than forced through, a filter, resulting in a pressure differential that keeps contaminants from re-mixing with newly purified samples.

Samples flow parallel to a filter and can be recycled repeatedly through a reservoir. Types of TFF include:

  • Micro filtration
  • Nano filtration
  • Ultra-filtration

Tangential Flow Filtration Advantages

There are many advantages to a tangential flow filtration system, including:

  • Ease of use: Pump, gauge and tubing set-up is simple (as in the picture below). Along with the sample in the reservoir, the system is fully operable
  • Speed of results: TFF operates continuously (rather than in batches) and rapidly separates and purifies biomolecules
  • Ability to multi-task: Lab workers can concentrate and diafilter samples on the same filtration system
  • Flexibility: TFF units can process samples as small as 10 ml and as large as several thousands of litres

When to Use a Tangential Flow Filtration System

TFF is best used for large-scale separation and filtration processes such as:

  • Protein isolation from culture supernatants
  • Recovering and purifying antibodies, plasmid DNA, or recombinant proteins
  • Extracting endotoxins from buffers, water and media solutions
  • Clarifying fermentation broths
  • Harvesting cells
  • Preparing chromatography samples
  • Fractionating dilute protein mixtures
  • Concentrating and desalting proteins and peptides
  • TFF can handle a large surface area and filter larger volumes without clogging

Masterflex Peristaltic Pump System for TFF

TFF in Action

TFF used in research of viruses.

  • TFF set-up using a Masterflex peristaltic pump system produced results with a higher concentration of viruses captured by the tangential flow filter than that from a system with less filtration control.
  • Case study: Study of the viruses that inhabit the intestines of a cow. To conduct this research, the researcher set up a system using a Masterflex® L/S® peristaltic pump and pump head with silicone tubing, pressure gauges, and fittings to mount gauges inline and to connect to the filter. They ran samples from a reservoir through the peristaltic pump and the pressure gauge into the tangential flow filter. They used two filters in separate processes: A 0.2 µm filter to remove bacteria and a <100 kD filter to concentrate the viruses.

TFF has been used by vintners and oenologists — winemakers and wine scientists —to:

  • Produce cleaner wines
  • Filter out cellular debris
  • Cut costs on bottling final filters

TFF has also found its way into food and pharmaceutical processing, where continuous purification speeds the innovation process.

Recommended TFF Equipment

Masterflex® recommends the following TFF equipment:

  • Masterflex® peristaltic digital process drives and pump heads — which meet all requirements and maintain sample integrity. Choose dual-channel pump heads so both filters can run simultaneously.
  • Masterflex® silicone tubing
  • Tangential flow filters
  • Pressure gauges
  • Fittings to mount gauges and connect the filter

Whatever your filtration needs, Masterflex® offers the latest, best quality solutions, always at scale.