UNOsphere S Media

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Overview

Type of ion exchanger	Strong cation
Functional group	-SO₃^-
Total ionic capacity	280 ± 30 µeq/ml
Dynamic binding capacity*
150 cm/hr	60 mg IgG/ml
1,200 cm/hr	30 mg IgG/ml
Median particle size	80 µm
Recommended linear flow rate range**	50–1,200 cm/hr
Chemical stability
1.0 M NaOH (20°C)	Up to 2,000 hr
1.0 M HCl (20°C)	Up to 200 hr
pH stability range	pH 1–14
Regeneration conditions	1–2 M NaCl
Storage conditions	20% Ethanol 0.1 N NaOH for 30 days

* 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.

Binding and Backpressure Properties

Binding and backpressure properties of UNOsphere S support. Column size, 1.1 x 20 cm; sample, 2 mg/ml human IgG; buffer, 50 mM sodium acetate, pH 5.0 (—), backpressure; (—), 10% breakthrough capacity.

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UNOsphere S Purification IgG

Purification of IgG

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.

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UNOsphere Media Purification Analysis

SDS-PAGE gel of UNOsphere-purified murine IgG1

SDS-PAGE gel of UNOsphere-purified murine IgG₁. Fractions from the chromatography run shown were separated on a 4–20% precast gel. Lane 1, conditioned medium; lane 2, flowthrough; lanes 3–8, fractions 25, 26, 27, 28, 29, and 30; lane 9, protein A–purified murine IgG₁ from culture medium.

UNOsphere S cation exchange media are designed to meet the increasing needs of the biopharmaceutical industry for high-productivity chromatographic media. UNOsphere, a hydrophilic polymeric support based on a single-step polymerization process, delivers high productivity through high binding capacity and low backpressures. The single-step polymerization process produces a support unsurpassed in quality and batch-to-batch reproducibility.

UNOsphere S support is a strong cation exchanger.

Features include:

Optimized to operate under 2 bar backpressure at linear velocities of 1,200 cm/hr
Stable to 1 M NaOH exposure up to 10,000 hours
Efficient capture of biomolecules from crude feedstreams
Large pores that result in ultrahigh binding capacities at fast linear velocities
Hydrophilic polymeric beads
Robust polymer designed to withstand repeated clean-in-place cycles
Biopharmaceutical manufacturing quantities available
Fully supported for regulatory submission

Most production chromatography systems have maximum pressure limits of 3 bar. The median particle size of UNOsphere S media is 80 µm, which generates a backpressure of less than 2 bar at 1,200 cm/hr. The highly macroporous nature of UNOsphere S media provides high binding capacities of 60 mg human IgG per ml of media at 150 ml/min. Harsh clean-in-place and sanitization cycles will not affect the long-term stability of UNOsphere media.

Regulatory support files are available upon request. Bio-Rad Laboratories is an ISO 9001-registered corporation.

Type of ion exchanger	Strong cation
Functional group	-SO₃^-
Total ionic capacity	280 ± 30 µeq/ml
Dynamic binding capacity*
150 cm/hr	60 mg IgG/ml
1,200 cm/hr	30 mg IgG/ml
Median particle size	80 µm
Recommended linear flow rate range**	50–1,200 cm/hr
Chemical stability
1.0 M NaOH (20°C)	Up to 2,000 hr
1.0 M HCl (20°C)	Up to 200 hr
pH stability range	pH 1–14
Regeneration conditions	1–2 M NaCl
Storage conditions	20% Ethanol 0.1 N NaOH for 30 days
* 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.