PARTICLES AND THEIR USE IN A METHOD FOR ISOLATING NUCLEIC ACID OR A METHOD FOR ISOLATING PHOSPHOPROTEINS
Monodisperse polymer microparticles comprising polystyrene or polyacrylate, wherein said particles have a coating formed of at least one transition metal oxide or porous polymer microparticles comprising polystyrene or polyacrylate, wherein said particles have a coating formed of at least one transition metal oxide. The use of such particles in a method for isolation of phosphoproteins from a sample containing phosphoproteins or for isolating nucleic acid from a sample containing nucleic acid is also described
1 . Polymer microparticles comprising polystyrene or polyacrylate, wherein said particles have a coating formed of at least one transition metal oxide of a group 4 or 5 metal or Al or a mixture thereof.
2 . Microparticles as claimed in claim 1 which are magnetic.
3 . Microparticles as claimed in claim 2 where the magnetic material is adsorbed onto the surface of the particles prior to the coating of the at least one transition metal oxide.
4 . Microparticles as claimed in claim 1 which are porous.
5 . Microparticles as claimed in claim 2 where the magnetic material is deposited in the pores of the particles prior to the coating of the at least one transition metal oxide.
6 . Microparticles as claimed in claim 1 which are covered by an additional polymeric coating layer prior to the coating of the at least one transition metal oxide.
7 . Microparticles as claimed in claim 1 which are monodisperse.
8 . Microparticles as claimed claim 1 having diameters of 0.2 to 10 μm.
9 . Microparticles as claimed in claim 1 wherein the transition metal oxide is titanium, tantalum, niobium or zirconium oxide.
10 . Microparticles as claimed in claim 1 being styrene divinylbenzene particles.
11 . A process for the preparation of microparticles as hereinbefore described comprising reacting polymeric microparticles comprising polystyrene or polyacrylate with at least one transition metal compound of a group 4 or 5 metal or Al or a mixture thereof capable of being converted into an oxide and forming the transition metal oxide coating, e.g. upon application of heat and/or water.
12 . A process as claimed in claim 11 wherein prior the microparticle starting material are surface functionalised.
13 . A process as claimed in claim 12 wherein the microparticles are surface functionalised to carry amino, hydroxyl, epoxide, carboxy or siloxy groups.
14 . A process as claimed in claim 11 wherein the transition metal compound is an alkoxide.
15 . A process as claimed in claim 14 wherein the alkoxide is a tetra(C1-6) zirconium or titanium alkoxide.
16 . A process as claimed in claim 11 carried out in at least two steps, a first step comprising contacting the microparticles with transition metal compound in an anhydrous environment and a second step comprising adding water to the product of the first step.
17 . A method for the isolation of phosphoproteins from a sample containing phosphoproteins comprising contacting said sample with the polymer microparticles of claim 1 .
18 . A method as claimed in claim 17 comprising:
(I) providing a sample that contains one or more phosphoproteins;
(II) mixing the sample with monodisperse polymer microparticles
(III) incubating the sample and the particles;
(IV) collecting the particles, optionally in a magnetic field and removing the supernatant; and optionally
(V) and eluting one or more isolated phosphoproteins.
19 . A method as claimed in claim 18 wherein step (V) is carried out in the presence of a buffer comprising phenyl phosphate and ammonium hydroxide.
20 . A method as claimed in claim 18 wherein step (II) is carried out in the presence of a buffer comprising water, ethanol and sodium acetate and having a pH of between 3.9 and 4.1.
21 . A method as claimed in claim 18 wherein step (II) is carried out in the absence of DHB.
22 . Use of the polymer microparticles as claimed in claim 1 in phosphoprotein isolation.
23 . A buffer comprising phenyl phosphate and ammonium hydroxide.
24 . A method for isolating nucleic acid from a sample containing nucleic acid comprising contacting said sample with polymer microparticles as claimed in claim 1 .
25 . A method as claimed in claim 24 in which contact between the nucleic acid and microparticles occurs in the presence of a chaotropic buffer.
26 . A method as claimed in claim 24 for the isolation of nucleic acid from a sample comprising:
(I) providing a sample that contains one or more nucleic acids;
(II) mixing the sample with transition metal oxide coated microparticles as hereinbefore described;
(III) incubating the sample and the particles;
(IV) collecting the particles, optionally in a magnetic field and removing the supernatant; and
(V) optionally eluting one or more isolated nucleic acids.
27 . Use of the polymer microparticles as claimed in claim 1 to isolate nucleic acid.
28 . A kit for the isolation of nucleic acids comprising polymer microparticles as described in claim 1 and a chaotropic buffer.