YMC Europe has launched Biopro IEX, a chromatographic material that is able to withstand the demands of clean-in-place (CIP) procedures in the pharmaceutical production process.
Cleaning in place is an important part of any pharmaceutical production process.
For this reason, normally all production equipment is designed in such a way that it can be cleaned completely and verifiably without the need to dismantle it.
The chromatographic media also have to comply with this requirement and have to be able to withstand the harsh conditions of the CIP cycles.
It is expected that after the CIP cycle they perform the same way they performed upon first usage.
Therefore the IEX material YMC Biopro SP 75um was tested with regards to its ability to withstand the CIP procedure.
As a result it was found that even after 20 CIP cycles the material showed the same values for dynamic binding capacity (DBC), recovery and separation that made it a high-class material in the first place.
During one cycle, first a chromatogram for the separation of three standard proteins was obtained.
Next, the DBC and recovery for Lysozyme were determined, before the medium was cleaned in place with 1M NaOH for 5 CV at a linear flow of 400cm/h.
The protein mixture used for the separation was made up of Ribonuclease A, Cytochrom C and Lysozyme.
These were separated at a flow rate of 180cm/h using a 20mM potassium phosphate buffer pH 6.8 as mobile phase A and 20mM potassium phosphate buffer pH 6.8 containing 0.5M NaCl as phase B.
The DBC for Lysozyme was determined at 10 per cent breakthrough.
For this, the column was loaded with Lysozyme in 20mM Glycin-NaOH buffer pH 9.0 at a linear flow of 800cm/h.
For the measurement of the recovery, 0.5M NaCl was added to the same buffer system (without Lysozyme).
The DBC and recoveries are plotted after each CIP step.
For every cycle the DBC stays at a good value of generally above 220mg Lysozyme per ml resin.
Also the recovery remains unchanged with generally 100 per cent.
There is no difference in the ability to separate the protein mixture between the various runs.
As a result it can be stated that the usage of 5 CV 1M NaOH at a flow rate of 400 cm/h is a suitable CIP-procedure.
Furthermore, the experiments show that the YMC Biopro SP 75um material withstands at least 20 such CIP cycles, while keeping its high-class properties for DBC, recovery and separation power.
YMC Biopro IEX materials are, therefore, well suited for usage in big biopharmaceutical downstream processes.
The resins are available as strong cation exchanger (YMC Biopro SP) and strong anion exchanger (YMC Biopro QA), both in particle size of 30um and 75um.
