Purification of substances from a biological sample
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
G01N-033/53
G01N-033/543
출원번호
US-0508138
(2000-10-13)
우선권정보
DE-0039218 (1997-09-08)
국제출원번호
PCT/EP98/05521
(1998-08-31)
국제공개번호
WO99/13330
(1999-03-18)
발명자
/ 주소
Hosel, Wolfgang
Lenz, Helmut
Peter, Jochen
출원인 / 주소
Roche Diagnostics GmbH
대리인 / 주소
Amick, Marilyn L.Roche Diagnostics Corporation
인용정보
피인용 횟수 :
20인용 특허 :
4
초록▼
The invention concerns a method for the purification of a target substance from a biological sample by immobilizing the target substance on a solid phase by means of a high affinity binding pair and subsequently eluting it by adding a partner of the binding pair in a free form. In addition reagent k
The invention concerns a method for the purification of a target substance from a biological sample by immobilizing the target substance on a solid phase by means of a high affinity binding pair and subsequently eluting it by adding a partner of the binding pair in a free form. In addition reagent kits for carrying out the method are disclosed.
대표청구항▼
1. A method for purifying a target substance from a sample comprising the steps of: (a) providing a solid phase with an immobilized reactant R1wherein R1comprises at least one group that binds specifically to said target and at least one group comprising a first partner of a binding pair, wherein
1. A method for purifying a target substance from a sample comprising the steps of: (a) providing a solid phase with an immobilized reactant R1wherein R1comprises at least one group that binds specifically to said target and at least one group comprising a first partner of a binding pair, wherein the binding pair is characterized by having an affinity constant from about 106l/mol to 1010l/mol, and wherein binding of R1to said solid phase occurs by means of an immobilized reactant R2which comprises a group comprising a second partner of said binding pair, (b) contacting said sample with said solid phase resulting in adsorption of said target onto said solid phase via binding to R1, (c) separating non-adsorbed sample components from said solid phase, (d) contacting said solid phase with a first partner of said binding pair or analog thereof in free form resulting in desorption of said target as a complex with R1from said solid phase, and (e) separating said complex from said solid phase. 2. The method of claim 1, wherein said target is selected from the group consisting of cells, cell fractions, cell organelles, viral particles, polypeptides, peptides, glycoproteins, lipoproteins, polysaccharides, nucleic acids, hormones, metabolites, neurotransmitters and mediators. 3. The method of claim 1, wherein said target is selected from the group consisting of polypeptides and nucleic acids. 4. The method of claim 1, wherein said solid phase is a particulate solid phase. 5. The method of claim 1, wherein said group R1is selected from the group consisting of antibodies and nucleic acids. 6. The method of claim 1, wherein said first partner is a hapten group and said second partner is an antibody directed against said hapten group. 7. The method of claim 6, wherein said first partner is selected from the group consisting of digoxigenin, fluorescein, dinitrophenol, thyroid hormones, ruthenium and rhodium-(bispyridyl)3complexes, theophylline, barbiturates and diphenylhydantoin. 8. The method of claim 1, wherein said first partner is a sugar and said second partner is a lectin. 9. The method of claim 6, wherein said first partner is a sugar containing a mannose group and said second partner is concanavalin A. 10. The method of claim 1, wherein said first partner is a receptor ligand having a maximum molecular weight of 4 kD and said second partner is a biological receptor. 11. The method of claim 1, wherein said first partner is a peptide epitope and said second partner is an antibody directed against said epitope. 12. The method of claim 1, wherein said desorption is carried out at a pH of 5 to 8. 13. The method of claim 1, wherein said first partner in free form is in molar excess relative to said bound first partner. 14. The method of claim 1, wherein said first partner in free form is a conjugate comprising a carrier molecule coupled to multiple molecules of said first partner. 15. The method of claim 1, wherein said first partner in free form is an analog of said bound first partner and said analog has a higher binding affinity for said second partner than said bound first partner. 16. The method of claim 1, additionally comprising the step of releasing said substance from said complex with R1. 17. The method of claim 1, wherein said reactant R1carries a detection group that is different from said first partner. 18. The method of claim 1, wherein said target substance is a recombinantly produced polypeptide. 19. The method of claim 18, wherein said recombinantly produced polypeptide is a fusion polypeptide comprising a first domain which binds specifically to said first partner and a second domain which binds specifically to said target. 20. The method of claim 9, wherein said polypeptide further comprises a protease cleavage site between said first and second domains.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (4)
Kaluza Brigitte (Habach DEX) Lenz Helmut (Tutzing DEX), Diagnostic test using chimeric antibodies.
Hoyle Nicholas R. (Tutzing DEX) Pappert Gunter (Tutzing DEX) Grol Michael (Feldafing DEX) Hubner-Parajsz Christa (Tutzing DEX), Method and monoclonal antibodies for vitamin B12 determination.
Lenz Helmut (Tutzing DEX) Kleinhammer Gerd (Tutzing DEX), Process for the determination of an immunologically-bindable substance involving a Fab fragment.
Childs, Ronald F.; Dey, Tapan K.; Filipe, Carlos; Ghosh, Raja; Kim, Marcus Y.; Komkova, Elena N.; Mika, Alicja M.; Zhou, Jinsheng, Absorbing biological substances from liquid with supported porous gels containing binding sites.
Childs, Ronald F.; Filipe, Carlos; Ghosh, Raja; Mika, Alicja M.; Zhou, Jinsheng; Komkova, Elena N.; Kim, Marcus Y.; Dey, Tapan K., Composite material comprising supported porous gel containing functional groups and method of separating substances.
Childs, Ronald F.; Filipe, Carlos; Ghosh, Raja; Zhou, Jinsheng; Komkova, Elena N.; Kim, Marcus Y.; Dey, Tapan K., Composite materials comprising supported porous gels.
Childs, Ronald F.; Filipe, Carlos; Ghosh, Raja; Mika, Alicja M.; Zhou, Jinsheng; Komkova, Elena N.; Kim, Marcus Y.; Dey, Tapan K., Composite materials comprising supported porous gels containing metal-affinity ligands.
Kanani, Dharmeshkumar M.; Sidhu, Navneet; Kachuik, Bradley A.; Honeyman, Charles H., Layered tubular membranes for chromatography, and methods of use thereof.
Kanani, Dharmeshkumar M.; Sidhu, Navneet; Kachuik, Bradley A.; Honeyman, Charles H., Layered tubular membranes for chromatography, and methods of use thereof.
Childs, Ronald F.; Filipe, Carlos; Ghosh, Raja; Mika, Alicja M.; Zhou, Jinsheng; Komkova, Elena N.; Kim, Marcus Y.; Dey, Tapan K., Method for separating a substance from a fluid.
Childs, Ronald F.; Filipe, Carlos; Ghosh, Raja; Mika, Alicja M.; Zhou, Jinsheng; Komkova, Elena N.; Kim, Marcus Y.; Dey, Tapan K., Separating substances with supported porous gels containing metal-affinity ligands complexed with metal ions.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.