Composite chromatographic sorbent of mineral oxide beads with hydroxyapatite-filled pores

a technology of mineral oxide beads and apatite, which is applied in the field of composite chromatographic sorbents of mineral oxide beads with hydroxyapatite-filled pores, can solve the problems of difficult to achieve with the use of prior-art adsorbents, and achieve high purity separation, superior properties, and high acuteness and precision separation

Inactive Publication Date: 2006-02-02
PALL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011] In accordance with the present invention, a chromatography sorbent is provided which combines the separation capabilities of hydroxyapatite crystals with the strength of a mineral oxide network. This composite sorbent exhibits superior properties when used in chromatographic separation, or in substance separation or development when used as the adsorbent packed or charged into a column or as a stationary phase agent in column chromatography. Chromatography using this sorbent as the adsorbent in batch separations or packed in either a fixed bed or fluidized bed column achieves high acuteness and precision separation and fractionation of substances having a minute difference in structure from one another. This was difficult to achieve with the use of prior-art adsorbents. Such substances may include biological macromolecule materials having a molecular weight of 104 to 109 Dalton, such as proteins, including immunoglobulins, interferon or enzymes, or nucleic acids, such as RNA, DNA or plasmids or viruses. The composite sorbent is indispensable for high purity separation and refining of a variety of ultimately useful substances obtained by gene recombination, cell fusion or cell culture en masse.
[0012] The composite chromatographic sorbent comprises mineral oxide beads with pores filled with apatite, particularly hydroxyapatite. The mineral oxide beads of the composite sorbent are characterized by high porosity, low surface area, high mean pore diameter, and high mechanical stability. Moreover, they show a density that facilitates packing of fixed-bed columns, increases the particle sedimentation velocity in batch, and permits the use of high velocity in fluidized-bed operations. The apatite crystals are protected by a very strong skeleton of mineral oxide, preferably zirconia.

Problems solved by technology

This was difficult to achieve with the use of prior-art adsorbents.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Zirconia Particles by Sol-Gel

[0057] A silica sol is prepared by mixing sequentially and progressively 150 ml of sodium silicate 35% with 200 ml of water and 100 ml of water and 100 ml of glacial acetic acid. Dry solid irregular zirconia powder (350 mg of 0.3 to 3 μm size) is dispersed in this suspension. Cerium oxide (10 g) and cerium nitrate (10 g) are then added under vigorous stirring. Under the above conditions the gelation process occurs at ambient temperature within 15 to 60 minutes.

[0058] After complete gelation, which takes a few hours, the gel is divided into small pieces by press-filtering it through a 200 μ sieve. The particles are suspended in clear water and recovered by filtration, washed and then dried at 80° C. under an air stream.

[0059] The silica gel that entraps the solid zirconia and ceria composite microparticles is progressively dehydrated. At this point, the particles are soft and show only very modest porosity. Then, the particles are fired ...

example 2

Preparation of Zirconia Particles by Suspension Polymerization

[0061] A silica sol is prepared by mixing sequentially and progressively 150 ml of sodium silicate 35% with 200 ml of water and 100 ml of water and 100 ml of glacial acetic acid. Dry solid irregular zirconia powder (350 mg of 0.3 to 3 μm size) is dispersed in this suspension. Cerium oxide (10 g) and cerium nitrate (10 g) are then added under vigorous stirring.

[0062] The resulting homogeneous suspension is slowly poured in an agitated paraffin oil bath containing 2% sorbitan sesquioleate and dispersed as small droplets. The suspension is heated at 80° C. while stirring. Under these conditions, the gelation process occurs at ambient temperature within 15 to 30 minutes.

[0063] The beads of a diameter ranging from 10 to 500 μm comprise a silica hydrogel trapped within its network solid microparticles of pre-formed zirconia and ceria. They are recovered by filtration, washed, and dried at 80° C. under an air stream. The gel ...

example 3

Preparation of Zirconia Beads by Spray Drying

[0064] A solution is prepared by mixing 231 g of zirconium nitrate and 143.6 g of yttrium nitrate in 1000 ml of distilled water. Yttrium oxide (144 g) and zirconia powder (752 g of 0.3 to 3 μm size) are added under gentle stirring to prevent the introduction of air bubbles.

[0065] The suspension is then injected into a vertical drying chamber through an atomization device, such as a revolving disk, a spray nozzle, or an ultrasonic nebulizer, together with a hot gas stream, preferably air or nitrogen. The hot gas stream causes rapid evaporation of water from the microdroplets. The gas is typically injected at 300-350° C. and exits the dryer at a temperature slightly above 100° C. Microparticles of original mineral oxides are consolidated into individual aggregates of spherical shape. Dry microbeads are then fired at a temperature close to the melting temperature of the zirconium oxide to irreversibly consolidate the network. After cooling...

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Abstract

A new adsorbent of a porous mineral oxide material with apatite crystals, preferably hydroxyapatite crystals, in the pores of the mineral oxide material is disclosed. The adsorbent is useful for protein and nucleic acid separations

Description

BACKGROUND OF THE INVENTION [0001] The present invention relates to a new adsorbent of a porous mineral oxide material with apatite crystals, preferably hydroxyapatite crystals, in the pores of the mineral oxide material. The adsorbent is useful for protein and nucleic acid separations. [0002] Apatite is a calcium phosphate material in crystalline form having the general formula Ca5(F, Cl, OH, ½ CO3) (PO4)3. One of the more common types of apatite is hydroxyapatite which has the formula [Ca2(PO4)2]3Ca(OH)2. It is useful as a packing material to be filled in columns for chromatographic separation of biopolymers, for example, proteins, enzymes, and nucleic acids. Its ability to adsorb such molecules depends on both the structure of the crystal itself and on the exposed surface area of the crystals. [0003] The technique for the preparation of hydroxyapatite utilizable for column chromatography was first developed by Tiselius et al. [Arch. Biochem. Biophys., 65:132-155 (1956)]. Hydroxya...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): B01D15/08B01J20/04B01J20/281B01J20/06B01J20/10B01J20/28B01J20/282B01J20/283B01J20/284B01J20/32C07K1/20G01N30/60G01N30/88
CPCB01J20/0211B01J20/28085B01J20/048B01J20/06B01J20/08B01J20/103B01J20/28078B01J20/28097B01J20/282B01J20/3234B01J20/3242B01J20/3268B01J2220/52B01J2220/58C07K1/20B01J20/286B01J20/3028B01J20/3064B01J20/3078B01J20/3204B01J20/3236B01J20/327B01J20/3272B01J20/3274B01J20/3282B01J20/3293B01J2220/42B01J20/3071B01J20/28004B01J20/0296B01J20/0292
Inventor BOSCHETTI, EGISTOGIROT, PIERRE
Owner PALL CORP
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