Blue Trisacryl® M Affinity Chromatography Sorbent by Pall Life Sciences Products - Biopharmaceutical Division

Introduction
BioSepra Blue Trisacryl® M is an affinity chromatographic sorbent used for the purification of a wide variety of enzymes and proteins such as kinases, albumin, interferons, and some coagulation factors. The basic matrix is Trisacryl GF2000, a macroporous non ionic sorbent on which Cibacron Blue is covalently immobilized. The initial matrix is a copolymer of N-acryloyl-2-amino-2 hydroxymethyl-1,3-propane-diol in bead form. Its macroporous structure allows a good diffusion and improves exchange kinetics.
Cibacron blue F3GA dye is strongly bound to the matrix through a six carbon spacer arm (Figure 1). The reaction is performed with the EEDQ coupling agent. This type of coupling prevents any leakage of the dye in normal working conditions.

Chemical and Mechanical Stability
The excellent chemical stability of Blue Trisacryl M is a function of the synthetic nature of Trisacryl sorbent and the enhanced stability of the ligand coupling mechanism.
The sorbent has an excellent mechanical stability. High flow rates with working pressures up to 3 bar can be used.

Experimental results have shown that the flow rate increases linearly with pressure up to about 40 cm per hour. However, the absolute linear flow rate and its relation to working pressure will vary according to the column height. Due to their extended lifetime, Blue Trisacryl M columns do not require frequent repacking, which is cost- and labour-saving. Experimental results showed no modification of the chromatographic properties of the sorbent after more than 100 cycles of albumin separation from whole plasma. On an industrial scale, Blue Trisacryl M was used over 700 cycles in the production of human albumin from IgG depleted plasma.

Capacity
The binding capacity of Blue Trisacryl M depends on the protein involved. Additionally, the capacity for a given protein may differ according to the animal species. For example, the binding capacity for bovine albumin is lower than for human albumin. Binding capacity can also vary according to the working conditions, especially pH. Due to its binding capacity and its outstanding flow rate, Blue Trisacryl M is more efficient than any commonly used sorbent. Its long lifetime and the absence of non-specific adsorption contribute to improve yields.

Presentation and Storage
Blue Trisacryl M is supplied as ready-to-use labpacks suspended in 1 M sodium chloride and 20% ethanol as bacteriostatic. Before the first use, the sorbent can be stored at room temperature.
Once opened, Blue Trisacryl sorbent or columns must be stored at +4 °C in a neutral buffer containing a bacteriostatic agent. They must never be frozen.