UNOsphere Q Media

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Overview

Binding and backpressure properties of UNOsphere Q support: A 5 mg/ml sample of BSA in 10 mMTris, pH8.5, was loaded onto a 1.1 x 20 cm column. Red, backpressure; blue, 10% breakthrough capacity.

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Purification and Analysis of Plasmid DNA

Purification of plasmid DNA on a UNOsphere Q column. Clarified bacterial lysate (10 ml, adjusted to pH 8.0) was loaded onto a 0.5 x 11 cm column (2.1 ml) in buffer A (10 mM sodium phosphate, 0.3 M NaCl, 1 mM EDTA, pH 8.0). The sample was eluted with a 0–40% gradient of buffer B (10 mM sodium phosphate, 1.0 M NaCl, 1 mM EDTA, pH 8.0) at a flow rate of 2 ml/min (600 cm/hr). The column was washed with 10 column volumes of 40% buffer B, followed by 100% buffer B for 5 column volumes. The effluent was monitored at 254 nm. Each fraction was 5 ml. Blue, A₂₅₄; red, conductivity; black, theoretical gradient.

Analysis of plasmid DNA purified on UNOsphere Q support. Fractions were separated on a 0.8% agarose gel. Lane 1, 1 kb marker (catalog #170-8204); lane 2, crude lysate; lane 3, flowthrough (fractions 2–6); lane 4, fractions 9–18; lane 5, fractions 19–30; lane 6, fractions 31–35; lane 7, fraction 36.

Restriction enzyme analysis of plasmid DNA purified on UNOsphere Q support. Lane 1, 1 kb marker; lane 2, undigested clarified lysate; lane 3, EcoRI-digested clarified lysate; lane 4, undigested fractions 31–35; lane 5, digested fractions 31–35; lane 6, undigested fraction 36; lane 7, digested fraction 36.

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UNOsphere ion 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 Q media are strong anion exchangers.

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 Q support is 120 µm, which generates a backpressure of less than 2 bar at 1,200 cm/hr. The highly macroporous nature of UNOsphere Q support provides high binding capacities that range from 180 to 125 mg BSA per ml in the linear velocity range of 150–1,200 cm/hr, respectively. Harsh clean-in-place and sanitization cycles will not affect the long-term stability or performance of UNOsphere Q media.

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

Type of ion exchanger	Strong anion
Functional group	-N(Ch₃)₃⁺
Total ionic capacity	120 µeq/m
Dynamic binding capacity*
at 150 cm/hr	180 mg BSA/ml
at 600 cm/hr	125 mg BSA/ml
Median particle size	120 µm
Recommended linear flow rate range**	50–1,200 cm/hr
Chemical stability
in 1.0 M NaOH (20°C)	Up to 10,000 hr
in 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 M NaOH for 30 days
* 10% breakthrough capacity determined with 2.0 mg/ml BSA in a 1.1 x 10 cm column.
** UNosphere Q support packed into a 20 cm bed height and run at 1,200 cm/hr generates less than 2 bar backpressure.