UNOsphere™ Cation Exchange Media

*  10% breakthrough capacity determined with 4.5 mg/ml human IgG in a 1.1 x 10 cm column.
** UNOsphere S support packed into a 20 cm bed height and run at 1,200 cm/hr generates less than 2 bar backpressure.

Purification of murine IgG1 on UNOsphere S column. Column size, 0.5 x 10 cm (2 ml); sample, 15 ml (6.6 mg) of murine IgG1-conditioned medium. The sample was loaded onto the column in 20 mM citrate-phosphate buffer, pH 4.0, washed, and eluted in a linear gradient of 0-100% 20 mM citrate-phosphate, pH 8.0 in 10 column volumes at a flow rate of 2.0 ml/min (600 cm/hr). Each fraction was 2.0 ml. (–), A₂₈₀; (–), buffer pH.

Stability of UNOsphere Rapid S media. Results from an accelerated storage study (in 0.1 M NaOH) show no loss in dynamic binding capacity in an equivalent-to-5-year test. DBC, dynamic binding capacity.

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An example of separation. A, chromatogram of IgG purification from calf serum (green trace) and purified bovine IgG (blue trace) using the the same binding and elution conditions. B, chromatography conditions.

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Purification of a Monoclonal Antibody Using a Combination of UNOsphere Rapid S and UNOsphere Q Media Ion Exchange Chromatography After Protein A Capture.

UNOsphere Rapid S media performance. Pressure/flow performance for a 20 cm diameter x 20 cm bed height column packed to 17% axial compression.

Dynamic binding capacity (DBC) vs. flow velocity. A column 1.1 cm in diameter was packed to a 20 cm bed height with UNOsphere Rapid S media; 2 mg/ml human IgG in 50 mM sodium acetate was loaded until 10% breakthrough was observed.

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