Hydroxyl group-containing covalently bound polymer separating ma

Details Hydroxyl group-containing covalently bound polymer separating ma Hydroxyl group-containing covalently bound polymer separating ma

1997-07-22

2000-10-24

Henderson, Christopher

Synthetic resins or natural rubbers -- part of the class 520 ser

Synthetic resins

Mixing of two or more solid polymers; mixing of solid...

525291, C08F 406, C08F 216

Patent

active

06136925&

DESCRIPTION:

BRIEF SUMMARY
SUMMARY OF THE INVENTION

The invention relates to separating materials based on supports containing hydroxyl groups, whose surfaces are coated with covalently bound polymers, and to processes for their preparation.
The separating materials according to the invention can be employed for the separation of macromolecules, in particular for the fractionation of biopolymers. The separation and purification of biological macromolecules, such as nucleic acids, proteins, enzymes, subcellular units, peptides, monoclonal antibodies or whole cells, has gained great importance with regard to genetic engineering and biotechnology.
For example, the use of ion exchangers for the fractionation of biological macromolecules is known. The conventional materials consist of polymers, such as polymethacrylates, polystyrenes, agarose, crosslinked dextran or silica gels, which carry appropriate functional groups.
EP 337, 144 discloses separating materials based on supports containing hydroxyl groups, whose surfaces are coated with covalently bound polymers, the polymers being identical or different recurring units which are bound to the support by graft polymerization in the presence of cerium(IV) ions.
As a whole, these separating materials are not optimal and, in particular with respect to the preparation process and the grafting yield, still have considerable disadvantages.
The invention is based on the object of making available an optimal separating material which does not have the disadvantages mentioned.
The invention relates to separating materials based on supports containing hydroxyl groups, whose surfaces are coated with covalently bound polymers, which are characterized in that the polymers consist of identical recurring units of the formula I
The invention further relates to processes for the preparation of these separating materials, which are characterized in that the graft polymerization is carried out in the presence of cerium(IV) ions and of 1 to 3.5 mol/l of inorganic salts in the polyacrylation mixture.
In this case, it is particularly advantageous if the monomers necessary for the polymerization are prepared by reaction of acrylate with aminoethanesulfonic acid in aqueous solution in the presence of a stabilizer and employed directly for the graft polymerization.
Surprisingly, it has been shown that the support materials according to the invention are particularly suitable for high-speed chromatographic separations. The separating materials are universally employable for the ion-exchange chromatography of macromolecules, in particular of biopolymers.
The separating materials according to the invention consist of support particles having hydroxyl groups, onto which is grafted a polymeric material via the .alpha.-C atoms of the hydroxyl groups, starting from the monomer sulfoethylacrylamide.
Possible support particles are all generally known porous and nonporous chromatographic supports which have primary or secondary, aliphatic hydroxyl functions on the surface.
Preference is given in this case, for example, to hydrophilic polymers based on acrylate and methacrylate, polymers based on polyvinyl alcohol, diol-substituted silica gels, polysaccharides based on agarose, cellulose, cellulose derivatives or polymers based on dextran. However, it is of course also possible to employ other polymers or copolymers based on monomers such as vinyl compounds, acrylamide, (meth)acrylic acid esters or (meth)acrylonitrile in hydroxylated form.
The performance of high-speed chromatographic separations in so-called downstream processing has recently acquired increasing importance. Two important aspects are in favor, for example in protein purification, of carrying out a high-speed separation: too long a contact of the protein to be purified with the support material leads to a decrease in the biological activity and the proteases released in cell disruption destroy the proteins during a long elution period.
An essential prerequisite for carrying out a high-speed separation, however, is that the protein binding capacity is inde

REFERENCES:
patent: 4547463 (1985-10-01), Sakata
patent: 4617321 (1986-10-01), MacDonald
patent: 5021160 (1991-06-01), Wolpert
EPA 0259037 Mar. 9, 1988.

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