DK2564203T3 - Antistof-nanopartikelkonjugater og fremgangsmåder til fremstilling og anvendelse af sådanne konjugater - Google Patents

Antistof-nanopartikelkonjugater og fremgangsmåder til fremstilling og anvendelse af sådanne konjugater Download PDF

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DK2564203T3
DK2564203T3 DK11721853.7T DK11721853T DK2564203T3 DK 2564203 T3 DK2564203 T3 DK 2564203T3 DK 11721853 T DK11721853 T DK 11721853T DK 2564203 T3 DK2564203 T3 DK 2564203T3
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antibody
sample
metal
nanoparticle
silver
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Julia Ashworth-Sharpe
Chol Steven Yun
Zhanna Zhilina
Adrian E Murillo
Donald D Johnson
Michael Farrell
Jerome W Kosmeder
Christopher Bieniarz
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Ventana Med Syst Inc
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    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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    • C07F9/02—Phosphorus compounds
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    • C07F9/08—Esters of oxyacids of phosphorus
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    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/655—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms
    • C07F9/6552—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a six-membered ring
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    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)

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Claims (22)

1. Fremgangsmåde til påvisning af et målmolekyle i en prøve, hvilken fremgangsmåde omfatter: etablering af kontakt mellem en prøve og et første antistof, der binder sig til målmolekylet; etablering af kontakt mellem prøven og et andet antistof, der er konjugeret til ét eller flere enzymmolekyler, hvor enzymmolekylerne er i stand til at danne mindst ét produkt, der er i stand til at reducere metalioner til metal i oxidationstilstand nul, og hvor det andet antistof specifikt binder sig til det første antistof; etablering af kontakt mellem prøven og et tredje antistof, hvor det tredje antistof er et antistof-nanopartikelkonjugat, der omfatter to eller flere nanopartikler, der er direkte bundet til et antistof via en metal-thiolbinding, og specifikt binder sig til det andet antistof; etablering af kontakt mellem prøven og et substrat for enzymmolekylerne og en metalion, således at et metalpræcipitat dannes og colokaliseres med målmolekylet; og påvisning af metalpræcipitatet, hvorved målmolekylet påvises.
2. Fremgangsmåde ifølge krav 1, hvor det andet antistof endvidere omfatter et hapten, og hvor det tredje antistof specifikt binder sig til det andet antistofs hapten.
3. Fremgangsmåde ifølge krav 1 eller 2, hvor målmolekylet omfatter en hapten-mærket nukleinsyreprobe, der specifikt binder sig til målmolekylet, og hvor det første antistof specifikt binder sig til den hapten-mærkede nukleinsyreprobe.
4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 3, hvor metalionen omfatter sølvion, guldion, kobberion, nikkelion, platinion, palladiumion, cobaltion eller iridiumion.
5. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 4, der endvidere omfatter etablering af kontakt mellem prøven og et guldhalogenidsalt, eventuelt hvor guldhalogenidsaltet omfatter guldchlorid.
6. Fremgangsmåde ifølge krav 5, hvor fremgangsmåden endvidere omfatter forstærkning af signalet, hvor forstærkning af signalet omfatter etablering af kontakt mellem prøven og et sølvsalt, eventuelt hvor sølvsaltet omfatter sølvnitrat, sølvoxid, sølvacetat eller sølvperchlorat.
7. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 6, der endvidere omfatter etablering af kontakt mellem prøven og et reduktionsmiddel, eventuelt hvor reduktionsmidlet omfatter natriumthiosulfat.
8. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 7, hvor det andet antistof omfatter et gede-antikanin-immunoglobulin G-antistof eller er konjugeret til tre alkaliske phosphatmolekyler.
9. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 8, hvor enzymet omfatter en alkalisk phosphatase, en sur phosphatase, en β-galactosidase, en β-glucosidase, en β-lactamase eller en esterase, eventuelt hvor enzymet er alkalisk phosphatase og substratet for enzymet omfatter 5-brom-4-chlor-3-indolylphosphat, ascorbinsyrephosphat eller et hyd roquinonphosphatderivat.
10. Fremgangsmåde til påvisning af et målmolekyle i en prøve, hvilken fremgangsmåde omfatter: etablering af kontakt mellem prøven og et første antistof, der er konjugeret til ét eller flere enzymmolekyler, hvor enzymmolekylerne er i stand til at danne mindst ét produkt, der er i stand til at reducere metalioner til metal i oxidationstilstand nul, og hvor det første antistof binder sig til målmolekylet; etablering af kontakt mellem prøven og et andet antistof, hvor det andet antistof er et antistof-nanopartikelkonjugat, der omfatter to eller flere nanopartikler, der er direkte bundet til et antistof via en metal-thiolbinding, og specifikt binder sig til det første antistof; etablering af kontakt mellem prøven og et substrat for enzymmolekylerne og en metalion, således at et metalpræcipitat dannes og colokaliseres med målmolekylet; og påvisning af metalpræcipitatet, hvorved målmolekylet påvises.
11. Fremgangsmåde ifølge krav 10, hvor det første antistof omfatter et hapten, og hvor det andet antistof er et anti-haptenantistof, der specifikt binder det første antistofs hapten.
12. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 11, hvor antistof-nanopartikelkonjugatet omfatter to til syv nanopartikler, eventuelt hvor konjugatet omfatter fem nanopartikler.
13. Fremgangsmåde ifølge krav 12, hvor metal-thiolbindingens metal er konjugeret til en cysteinrest hos antistoffet.
14. Fremgangsmåde ifølge krav 12 eller 13, hvor antistoffet hos antistof-nanopartikelkonjugatet omfatter et antistof eller antistoffragment.
15. Fremgangsmåde ifølge et hvilket som helst af kravene 12 til 14, hvor antistof-nanopartikelkonjugatets antistof omfatter et kanin-, gede-muse- eller anti-hapten-antistof.
16. Fremgangsmåde ifølge et hvilket som helst af kravene 12 til 15, hvor nanopartiklerne omfatter guld, palladium, platin, sølv, kobber, nikkel, cobalt, iridium eller en legering af to eller flere deraf.
17. Fremgangsmåde ifølge et hvilket som helst af kravene 12 til 16, hvor nanopartiklen er 5 nm eller mindre i diameter, eventuelt hvor nanopartiklen er 0,5 til 5 nm i diameter.
18. Kit til påvisning af et målmolekyle i en prøve, hvilket kit omfatter én eller flere beholdere, der omfatter: et første antistof, hvor det første antistof er et antistof-nanopartikelkonjugat, der omfatter to eller flere nanopartikler, der er direkte bundet til et antistof via en metal-thiolbinding; et andet antistof, der er konjugeret til ét eller flere enzymmolekyler, hvor enzymmolekylerne er i stand til at danne mindst ét produkt, der er i stand til at reducere metalioner til metal i oxidationstilstand nul, og hvor det første antistof specifikt binder sig til det andet antistof; og et substrat for det ene eller flere enzymmolekyler og en metalion.
19. Kit ifølge krav 18, der endvidere omfatter et guldhalogenid, eventuelt hvor guldhalogenidet omfatter guldchlorid.
20. Kit ifølge krav 18 eller 19, der endvidere omfatter et sølvsalt, eventuelt hvor sølvsaltet omfatter sølvnitrat, sølvoxid eller sølvchlorid.
21. Kit ifølge et hvilket som helst af kravene 18 til 20, der endvidere omfatter et reduktionsmiddel, eventuelt hvor reduktionsmidlet omfatter natriumthiosulfat.
22. Kit ifølge et hvilket som helst af kravene 18 til 21, der endvidere omfatter et tredje antistof, der specifikt binder et målmolekyle, hvor det andet antistof specifikt binder sig til det tredje antistof.
DK11721853.7T 2010-04-27 2011-04-27 Antistof-nanopartikelkonjugater og fremgangsmåder til fremstilling og anvendelse af sådanne konjugater DK2564203T3 (da)

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