Polymeric adsorbents and method of preparation

Abstract

Macroporous polymers having selected porosity and permeability characteristics that provide rigid polymer matrices suitable for use in medium and high pressure reversed phase liquid chromatography (RPC) are disclosed. A method for preparing the polymers using selected mixed porogens in selected proportions relative to the monomer phase is also disclosed. The polymers are especially useful as stationary phases in large scale chromatography columns without developing increased pressures during prolonged use, while maintaining good chromatographic performance for targeted biomolecules, such as insulin.

Description

[0001] This invention relates to novel macroporous polymers having selected porosity and permeability characteristics that provide rigid polymer matrices suitable for use in medium and high pressure reversed phase liquid chromatography (RPC). The polymers are especially useful as stationary phases in large scale chromatography columns without developing increased pressures during prolonged use.[0002] Stationary phases useful in RPC, especially in high performance preparative mode (such as required in the separation and purification of biomolecules), must be mechanically rigid to withstand the high operating pressures generated within the chromatography columns. Silica matrices, which have been commonly used for these applications in the past, have satisfactory mechanical rigidity; however, silica matrices cannot be operated under high pH conditions, which severely limits their use in a wide range of biomolecule separations. This factor limits the purification options available to th...

AI Technical Summary

Benefits of technology

[0011] We have discovered that novel macroporous polymers useful for large scale separation and purification of biomolecules by high pressure liquid reverse phase chromatography can be prepared having selected porosity and permeability characteristics. In particular, we have discovered that using specific porogen solvents in specific proportions relative to the monomer phase under specific polymerization conditions unexpectedly provides the rigid polymer matrices of the present invention. The novel macroporous polymers may be used in high pressure RPC without significant compressibility and pressure buildup while maintaining good throughput and capacities (dynamic and equilibrium) for targeted biomolecules.

[0052] The macroporous polymers of the present invention typically have a total porosity of 0.7 to 2, preferably from 0.9 to 1.8 and more preferably from 1.0 to 1.7 cm.sup.3 / g. Typically, the macroporous polymers have an operational mesoporosity of 0.7 to 1.9, preferably from 0.8 to 1.7 and more preferably 0.9 to 1.4 cm.sup.3 / g. Typically, the macroporous polymers have an operational microporosity from zero to 0.5, preferably from zero to 0.3, more preferably from zero to 0.2 and most preferably from zero to less than 0.1 cm.sup.3 / g. Typically, the macroporous polymers have a macroporosity from zero to 0.6, preferably from zero to 0.5 and more preferably from zero to 0.3 cm.sup.3 / g. Macroporosity values above about 0.6 cm.sup.3 / g decrease the working capacity of the polymer for biomolecules of the targeted molecular size and shape, in terms of total capacity.

Problems solved by technology

Silica matrices, which have been commonly used for these applications in the past, have satisfactory mechanical rigidity; however, silica matrices cannot be operated under high pH conditions, which severely limits their use in a wide range of biomolecule separations.

This factor limits the purification options available to the process chromatographer, and adversely affects the production lifetime of silica media (they degrade faster because they cannot be cleaned under aggressive conditions), resulting in poorer overall economics of commercial manufacturing processes.

However, current polymeric stationary phases are somewhat compressible at the medium to high pressure conditions used in high-performance biomolecule separations.

This compressibility is detrimental to separation processes because it limits the range of operable flowrates, and it can degrade the integrity of the polymer bed in the column.

For example, the following references disclose polymers used at column conditions representative of high-pressure analytical operations (less than 0.5 cm internal diameter columns), where "wall effects" are known to minimize polymer compressibility; however, these references do not disclose operations in larger scale, high-pressure commercial chromatography columns where one would expect additional pressure buildup from polymer compressibility due to the absence of wall effects: Lloyd, L. L. and Warner, F. P., Preparative High Performance Liquid Chromatography on a Unique High-Speed Macroporous Resin, J. Chromatography, Vol. 512, pp 365-376 (1990); and Lloyd, L. L., Rigid Macroporous Copolymers as Stationary Phases in High Performance Liquid Chromatography, Review, J. Chromatography, Vol. 544, pp 201-217 (1991).