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Tutorials : Apr 1, 2008 ( )
Removing Color Impurities in Biomanufacturing
Using Inexpensive Resins Upstream Improves Purification and Downstream-Column Life
Within the pharmaceutical and biopharmaceutical industry, there are many process streams that require purification by some means or another. Typically, this requires that a specific impurity is removed or that a range of impurities are eliminated. In the latter case, these impurities are never fully characterized and are generally called color-species.
As mentioned earlier, most degradation products tend to be polymeric species containing one or several carboxylic acid groups. These colors are inclined to be large organic molecules that have a weakly negative charge. The types of resins that are typically used in these cases are ion-exchange resins that have a defined pore structure (macroreticular) or gel matrix and a positive charge derived from either a quaternary amine or tertiary amine function.
The Table includes a list of ion exchange resins that are widely used for decolorization, but other similar products are available.
A filtered, crude vancomycin broth was adjusted to pH 8, and the broth was then pumped over a column of Amberlite FPA40 Cl using a Rainin Rabbit Pump. The column effluent was monitored with a Linear Model 200 detector (Grace Davison Discovery Science) and fractions were collected in one tube every five minutes using a Gilson 201 Fraction Collector. The experiment involved a column of 2.0 cm ID x 30 cm L and a flow rate of 1.63 mL/minute (1 CV/Hr). Detection was performed with a Thermo Fisher Scientific Biomate 3 Spectrophotometer at 400 nm.
The broth was pumped over the column for a total of five column volumes (470 mL) before the experiment was terminated and then reactions of the effluent were analyzed.
The average color removal was 50% (Figure 1) after the first column volume had passed through the column. Figure 2 shows a close-up of Rohm & Haas’ Amberlite™ FPA40 Cl column with the color adsorbed on the top of the column.
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