Method for isolating proteins or protein and nucleic acid associations, or particle and protein complexes, reagent and uses
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
A23J-001/00
C07K-001/00
C07K-014/00
C07K-016/00
C07K-017/00
출원번호
US-0182126
(2001-01-22)
우선권정보
FR-00 00862(2000-01-21)
국제출원번호
PCT/FR01/000205
(2001-01-22)
§371/§102 date
20030113
(20030113)
국제공개번호
WO01/052612
(2001-07-26)
발명자
/ 주소
Elaissari,Abdelhamid
Mandrand,Bernard
Delair,Thierry
Spencer,Doran
Arkis,Ahmed
출원인 / 주소
Biomerieux
대리인 / 주소
Oliff &
인용정보
피인용 횟수 :
5인용 특허 :
7
초록▼
The invention concerns a method for isolating proteins and/or protein and nucleic acid associations in a sample, comprising steps which consist in: contacting said sample and magnetic colloidal particles comprising a core and a coat wherein: the core is magnetic and is coated with at least a polymer
The invention concerns a method for isolating proteins and/or protein and nucleic acid associations in a sample, comprising steps which consist in: contacting said sample and magnetic colloidal particles comprising a core and a coat wherein: the core is magnetic and is coated with at least a polymer comprising functional groups X selected among amine, hydroxyl, thiol, aldehyde, ester, anhydride, acid chloride, carbonate, carbamate, isocyanate and isothiocyanate groups or mixtures thereof, whereof at least one fraction has reacted with other functional groups of the coat, and the coat consists of a polymer bearing functional groups Z and Z', capable of ionisation, identical or different, selected among amine, carboxylic acid, ester, anhydride, aldehyde, thiol, disulphide, α-halogenocarbonyl, sulphonic acid, maleimide, isocyanate and isothiocyanate groups to form a mixture; incubating said mixture in predetermined conditions; and separating from the mixture the protein and/or protein and nucleic acid associations complexed on the colloidal particles by applying a magnetic field. The invention also concerns a complex consisting of colloidal particles and proteins, a reagent comprising such a complex or colloidal particles.
대표청구항▼
The invention claimed is: 1. A method for isolating at least one of proteins and protein and nucleic acid associations, from a sample, comprising: bringing said sample into contact with magnetic colloidal particles to form a mixture, said magnetic colloidal particles comprising a core and an envelo
The invention claimed is: 1. A method for isolating at least one of proteins and protein and nucleic acid associations, from a sample, comprising: bringing said sample into contact with magnetic colloidal particles to form a mixture, said magnetic colloidal particles comprising a core and an envelope in which: the core is magnetic and is coated with at least one polymer comprising functional groups X chosen from amine, hydroxyl, thiol, aldehyde, ester, anhydride, acid chloride, carbonate, carbamate, isocyanate and isothiocyanate groups, or mixtures thereof, at least one fraction of which has reacted with other functional groups of the envelope, and the envelope comprises a polymer bearing ionizable functional groups, Z and Z', which may be identical or different, chosen from amine, carboxylic acid, ester, anhydride, aldehyde, thiol, disulfide, α-halocarbonyl, sulfonic acid, maleimide, isocyanate and isothiocyanate groups, which have partially reacted with the functional groups X of the core; incubating said mixture under predetermined conditions, and applying a magnetic field to separate at least one of proteins and protein and nucleic acid associations complexed on said colloidal particles from the mixture. 2. The method as claimed in claim 1, wherein the core comprises: at least one organic polymer chosen from at least one homopolymer or at least one copolymer, or mixtures thereof, derived from polymerization of at least one monomer chosen from monomers of acrylamide, monomers of acrylate, styrene, methylstyrene, ethylstyrene, tert-butylstyrene, chloromethylstyrene, vinyltoluene; derivatives thereof and copolymers of these monomers with one another, with other comonomers or with one another and other comonomers, and metal oxide particles chosen from particles of iron oxide, titanium oxide, cobalt oxide, zinc oxide, copper oxide, manganese oxide, nickel oxide; magnetite; hematite, ferrites, cobalt oxide alloys, and nickel oxide alloys. 3. The method as claimed in claim 1, wherein the polymer of said envelope is chosen from at least one hydrophilic homopolymer or copolymer chosen from: homopolymers or copolymers derived from polymerization of at least one monomer chosen from monomers derived from acrylamide or from methacrylamide; acrylic acid, methacrylic acid; acrylate derivatives, methacrylate derivatives; allylamine; styrene derivatives; on the condition, if the polymer is a homopolymer, the homopolymer comprises ionizable functional groups, polysaccharides, polypeptides, linear or branched polyethyleneimine, and dendrimers. 4. The method as claimed in claim 3, wherein the polymer of the envelope is poly(maleic anhydride vinyl ether), poly(N-vinylmorpholine-N-acryloxysuccinimide) or poly(N-vinylpyrrolidone-N-acryloxysuccinimide). 5. The method as claimed in claim 1, wherein the mixture is subjected to incubation at a temperature of between 15 and 60째 C. for an incubation time of between 5 and 60 minutes. 6. The method as claimed in claim 1, wherein the sample is a specimen or a culture supenatant comprising at least one of proteins and protein and nucleic acid associations. 7. The method as claimed in claim 6, wherein the sample is a biological sample. 8. A method for extracting at least one biological material selected from the group consisting of proteins and nucleic acids, comprising: isolating at least one member selected from the group consisting of viruses, bacteria, yeast, cells and mixtures thereof from a sample, according to the method described in claim 1, and if necessary for the extraction of the at least one biological material, subjecting the at least one separated member to at least one of a step of partial or total release and a step of denaturation to extract the at least one biological material from the at least one separated member. 9. The method as claimed in claim 2, wherein said monomers of acrylamide are at least one of N-alkylacrylamides and N,N-dialkylacrylamides. 10. The method as claimed in claim 9, wherein said monomers of acrylamide are selected from the group consisting of N-isopropylacrylamide, N-methylacrylamide, N-ethylmethacrylamide, N-n-propylacrylamide, N-n-propylmethacrylamide, N-isopropylmethacrylamide, N-cyclopropylacrylamide, N,N-diethylacrylamide, N-methyl-N-isopropylacrylamide or N-methyl-N-n-propylacrylamide. 11. The method as claimed in claim 2, wherein said monomers of acrylate are at least one of alkylacrylates and alkylmethacrylates in which the alkyl group comprises from 3 to 20 carbon atoms. 12. The method as claimed in claim 2, wherein said ferrites are selected from the group consisting of manganese ferrite, nickel ferrite and manganese-zinc ferrite. 13. The method as claimed in claim 3, wherein the at least one hydrophilic homopolymer or copolymer comprises at least one of copolymers or homopolymers of maleic anhydride and homopolymers or copolymers of acryloxysuccinimide. 14. The method as claimed in claim 3, wherein said polysaccharides are at least one of chitosan and poly(galacturonic acid). 15. The method as claimed in claim 3, wherein said polypeptides are at least one of polylysine and polyarginine. 16. The method as claimed in claim 5, wherein said temperature is between 20 and 35째 C. 17. The method as claimed in claim 5, wherein said incubation time is about 10 minutes. 18. The method as claimed in claim 6, wherein the sample comprises at least one of viruses, bacteria, yeast and cells. 19. The method as claimed in claim 7, wherein said biological sample is selected from the group consisting of a tissue specimen, a specimen of whole blood, a specimen of plasma, a specimen of serum, a culture supenatant and a agrofoods specimen. 20. The method as claimed in claim 8, wherein said sample is a specimen or culture supematant. 21. The method as claimed in claim 1, wherein during said incubating at least one of proteins and protein and nucleic acid associations react lonically with ionizable functional groups of said envelope and are thereby complexed on said colloidal particles. 22. A reagent for at least one of extracting, identifying, detecting and quantifying at least one of proteins and protein and nucleic acid associations, said reagent comprising colloidal particles comprising a core and an envelope in which: the core is magnetic and is coated with at least one polymer comprising functional groups X chosen from amine, hydroxyl, thiol, aldehyde, ester, anhydride, acid chloride, carbonate, carbamate, isocyanate and isothiocyanate groups, or mixtures thereof, at least one fraction of which has reacted with other functional groups of the envelope, and the envelope comprises a polymer bearing ionizable functional groups, Z and Z' , which may be identical or different, chosen from amine, carboxylic acid, ester, anhydride, aldehyde, thiol, disulfide, α-halocarbonyl, sulfonic acid, maleimide, isocyanate and isothiocyanate groups, which have partially reacted with the functional groups X of the core. 23. The reagent as claimed in claim 22, further comprising at least one means for at least one of identifying, detecting and quantifying said proteins or said nucleic acids. 24. A pharmaceutically acceptable vehicle for a vaccinal composition, characterized in that it consists of a particle comprising a core and an envelope in which: the core is magnetic and is coated with at least one polymer comprising functional groups X chosen from amine, hydroxyl, thiol, aldehyde, ester, anhydride, acid chloride, carbonate, carbamate, isocyanate and isothiocyanate groups, or mixtures thereof, at least one fraction of which has reacted with other functional groups of the envelope, and the envelope comprises a polymer bearing ionizable functional groups, Z and Z', which may be identical or different, chosen from amine, carboxylic acid, ester, anhydride, aldehyde, thiol, disulfide, α-halocarbonyl, sulfonic acid, maleimide, isocyanate and isothiocyanate groups, which have partially reacted with the functional groups X of the core. 25. The reagent according to claim 23, wherein said at least one means comprises at least one of a primer and a probe specific for nucleic acid of said protein and nucleic acid association.
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