PROCESS FOR MANUFACTURING A COMPOSITE SORBENT MATERIAL FOR CHROMATOGRAPHICAL SEPARATION OF BIOPOLYMERS
The present invention relates to a sorbent material for separation and purification of biopolymers, particularly nucleic acids, having a solid support substantially modified with a copolymer coating comprising aromatic monomers and crosslinking compounds and unsaturated esters or ethers preferably attached to the support via a vinylchlorsilane. The use of these materials for separation of nucleic acids, particularly a one-step isolation of DNA from lysates of different biological sources, is an object of the invention as well as a chromatographic column or cartridge at least partially filled with the sorbent material of the invention, a membrane-like device comprising the sorbent material of the invention, and a kit comprising the sorbent material of the invention in bulk or packed in chromatographic devices as well as other devices necessary for performing sample preparations.
1 - 22 . (canceled)
23 . A process for the purification and/or isolation of nucleic acids wherein the process comprises
lysing cells yielding a sample containing nucleic acids and proteins,
contacting the sample with a sorbent material binding the proteins and
collecting the eluate containing the nucleic acids
with the proviso that the sorbent material comprises a support which is at least partially covered by a polymer coating having the general formula
wherein x, y are, independent of each other, an integer of 1-100 and z is an integer of 0-100,
obtainable by a process comprising the steps of
modifying the support with a vinylchlorosilane
followed by addition of
a) at least one unsubstituted or substituted styrene or vinylnaphthalene [A] of general formula
(b) at least one cross-linkable unsubstituted or substituted (1, 4 or 1,2)-divinylbenzene [B] of general formula
c) optionally a non-saturated carboxylic acid or alkyl, aryl or hydroxylalkyl ester thereof [M] of general formula
or a non-saturated alcohol or ether of general formula
polymerising the copolymer composition
removing unreacted material and
recovering and drying the sorbent material.
24 . The process of claim 23 wherein the support is a porous inorganic material comprising inorganic metal oxide.
25 . The process of claim 24 wherein the porous inorganic metal oxides show a bidisperse distribution of pore sizes.
26 . The process of claim 23 wherein the support has an average pore size of 2-200 nm.
27 . The process of claim 23 wherein the polymer coating has a thickness of about 10 to 250 Angstrom.
28 . The process of claim 23 wherein the mass ratio of copolymer components [A], [B], and [M] complies with the formula [A]:[B]:[M]=1:0.03-0.30:0-0.2.
29 . The process of claim 24 wherein the inorganic metal oxide is selected from the group consisting of oxides of aluminum, titanium, zirconium, silicon oxides, iron oxides, controlled pore glass (CPG), diatomaceous earth and combinations thereof.
30 . The process of claim 25 wherein the porous inorganic metal oxide shows a bidisperse distribution with mean pore diameters in the range of 20-100 nm for the larger pore size.
31 . The process of claim 30 wherein the porous inorganic metal oxide has a mean pore diameter in the range of 2-15 nm for the smaller pore size.
32 . The process of claim 30 wherein the ratio of the mean diameter of the large pore size distribution and the lower pore size distribution is in the range of 2-15.
33 . The process of claim 26 , wherein the support has an average pore size of 2-100 nm.
34 . The process of claim 26 wherein the support has a specific surface area of 20-300 m 2 /g.
35 . The process of claim 34 wherein the support has a specific surface area of 20-100 m 2 /g.
36 . The process of claim 27 wherein the polymer coating has a thickness of 10 to 100 Angstrom.
37 . The process of claim 27 wherein the polymer coating has micropores of less than 50 Angstrom accessible to water, salts, and low molecular weight substances.
38 . The process of claim 27 wherein the polymer coating is non-adsorptive towards nucleic acids and adsorptive towards proteins.
39 . The process of claim 23 comprising component [M].
40 . The process of claim 39 , wherein [M] is selected from the group consisting of hydroxyethyl methacrylate, methacrylic acid and allyl alcohol.
41 . A process for the purification and/or isolation of nucleic acids wherein the process comprises
lysing cells yielding a sample containing nucleic acids and proteins,
contacting the sample with a sorbent material binding the proteins, and
collecting the eluate containing the nucleic acids
where the sorbent material comprises a support which is at least partially covered by a polymer coating having the general formula
wherein the mass ratio of copolymer components [A], [B], and [M] complies with the formula 1:0.03-0.3:0-0.2 and
[A] is derived from a monomer with general formula
[B] is derived from a monomer of general formula
and
[M] is derived from a monomer of general formula
or a monomer of general formula