APPLICATION NOTE


49


Purification of Monoclonal Antibodies with BioPro SmartSep


by Michael Ostendorf, YMC Europe GmbH • Schöttmannshof 19 • 46539 Dinslaken, Germany • Email: info@ymc.de


Ion exchange (IEX) is widely used for the analysis and purification of bio-molecules such as proteins, peptides and monoclonal antibodies. In industrial-scale production, IEX is used for initial capture, intermediate purifications or final product polishing.


Challenges During mAb Purification


For purification of monoclonal antibodies, high demands are required from the separating material. Factors influencing the binding characteristics of IgG are pH, linear velocity and/or salt concentration (conductivity) at the time the sample is loaded onto the column.


Therefore, a material with highly stable performance is required with regard to all those factors. In order to demonstrate the performance of YMC-BioPro materials, several studies have been performed.


• Purification of Adalimumab • Influence of pH


• Influence of linear velocity • Influence salt concentration


IgG Purification Scheme for Adalimumab


Column: Eluent:


Gradient: Flow rate:


50 x 5.0 mm ID


A) 20 mM citric acid-NaOH (pH 5.3) B) Eluent A containing 0.5 M NaCl


0-100% B (0-30 CV) 180 cm/hr (0.59 mL/min)


Temperature: ambient Detection: Sample:


lgG Load: Injection:


UV at 280 nm Anti-hTNFalpha


(after affinity chromatography) 0.1 mg 0.25 mL


In order to demonstrate the behaviour of BioPro SmartSep under different elution condi¬tions, experiments with different values of pH, linear velocity and salt concentration were performed and the dynamic binding capacity (DBC) recorded.


The parameters changed were:


Experimental Conditions pH: 6.0 vs. 5.3 Linear velocity: 200 - 800 cm/hr Salt concentration: 0 - 50 mM NaCl


-- Tosoh TSKgel SP-3PW -- GE Source 30S -- BioPro Smart Sep S30


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