Protein purification technology has been developed from a variety of methods such as chemical precipitation sample concentration or dialysis buffer exchange to the use of ultrafiltration membrane pressure-driven purification cross-flow systems. Ultrafiltration technology relies on the use of polymer membranes with highly defined pore sizes to separate molecules based on size. Ultrafiltration separation is based on mechanical interactions rather than chemical interactions, allowing researchers to concentrate samples without adding denaturing solvents or salts. Tangential flow filtration (TFF) is an ultrafiltration mode of operation. Alfa Chemistry explores tangential flow filtration and provides you with excellent laboratory TFF solutions.
About Tangential Flow Filtration
TFF is a fast and effective method for the separation and purification of biomolecules. It can be applied to a wide range of biological fields such as immunology, protein chemistry, molecular biology, biochemistry, microbiology, etc. The main applications of TFF are concentration, diafiltration (desalting and buffer exchange), and fractionation of large and small biomolecules. In addition, it can also be used to clarify and remove cells and cell debris in fermentation or cell culture fluids.
Figure.1 Process flow diagram for capture and purification of pDNA using tangential flow filtration and frontal anion-exchange membrane chromatography. (Guerrero-Germán, et al. 2009)
The following are the advantages of using tangential flow filtration in the laboratory:
- Easy to set up and use - Simply connect the TFF device to a pump and pressure gauge with tubing and some accessories, add the sample to the reservoir, and start filtering.
- Perform two steps with one system - Concentrate and percolate samples on the same system, saving time and avoiding product loss.
- Fast and efficient - It is easier to set up and faster than dialysis. Compared with the use of a centrifugal device or a stirred tank, a higher concentration can be reached in a shorter time.
- Enlarge or reduce proportionally - The materials of construction and box path length allow the conditions established during pilot-scale trials to be applied to process scale applications.
- economy - TFF devices and cassettes can be cleaned and reused, or discarded after a single-use.
Laboratory Tangential Flow Filter System
We provide a complete laboratory-scale TFF system. The system uses a plug-and-play design. Insert the TFF capsule, add the sample, and then turn on the pump to start processing. The TFF system includes all the hardware, piping, and accessories needed to get your TFF process up and running quickly.
The TFF capsule contains a polyethersulfone membrane with an effective filtration area of 50 cm2. Ultrafiltration membranes can provide a variety of molecular weight cut-offs, have low adsorption characteristics, and can achieve high product recovery rates. Sample batches of up to 1 liter can be desalted and then concentrated to a volume as low as 5 mL, requiring almost no user intervention. Reusable/disposable capsules can use the same equipment in a closed-loop connection to perform a single or continuous concentration/diafiltration step, saving time and avoiding product loss associated with the transfer step.
Scalable Tangential Flow Filter System
For larger and scalable applications, we provide a series of T series TFF cassettes for TFF systems, using superior structural materials and improved cassette design to improve process safety, reliability, reproducibility, and productivity.
- The feed and permeable screen material of the cassette is made of polypropylene, which is very resistant to sodium hydroxide.
- Larger feed ports and permeate ports can provide lower pressure drops.
- The T-series bellows are designed to provide maximum mass transfer through the membrane, which can shorten processing time or reduce installation area compared to existing similar bellows formats.
- The -series membrane packs contain our polyether sulfone membranes, which provide high throughput and selectivity. They have been specially modified to minimize the binding of proteins to membrane surfaces and interstitial structures.
Reference
- Guerrero-Germán P, et al. (2009). “Purification of Plasmid DNA Using Tangential Flow Filtration and Tandem Anion-exchange Membrane Chromatography.” Bioprocess and Biosystems Engineering. 32: 615-623.