Stimuli-responsive polymer diagnostic assay comprising magnetic nanoparticles and capture conjugates
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
G01N-033/543
B01D-021/00
G01N-001/34
B82Y-025/00
C12N-015/10
G01N-027/74
C12Q-001/68
B03C-001/01
B03C-001/28
G01N-001/40
출원번호
US-0732428
(2015-06-05)
등록번호
US-9429570
(2016-08-30)
발명자
/ 주소
Stayton, Patrick S.
Lai, Jriuan
Nehilla, Barrett J.
Srinivasan, Selvi
출원인 / 주소
University of Washington through its Center for Commercialization
대리인 / 주소
Christensen O'Connor Johnson Kindness PLLC
인용정보
피인용 횟수 :
0인용 특허 :
55
초록
The present disclosure utilizes the aggregation of stimuli-responsive polymers to isolate a diagnostic target (e.g., an antigen) from a solution using magnetophoresis. Isolating the diagnostic target provides a route to identify the presence of the diagnostic target in the solution.
대표청구항▼
1. A method of concentrating a target in a liquid, comprising applying a magnetic field to an aggregate in the liquid to provide a collected aggregate by magnetophoresis, wherein the aggregate comprises: (a) a stimuli-responsive magnetic nanoparticle comprising a first stimuli-responsive polymer att
1. A method of concentrating a target in a liquid, comprising applying a magnetic field to an aggregate in the liquid to provide a collected aggregate by magnetophoresis, wherein the aggregate comprises: (a) a stimuli-responsive magnetic nanoparticle comprising a first stimuli-responsive polymer attached to a magnetic core; and(b) a stimuli-responsive capture conjugate comprising a second stimuli-responsive polymer attached to a capture moiety, wherein the capture moiety is capable of binding to a target;wherein the aggregate is formed through associative interaction between the first stimuli-responsive polymer and the second stimuli-responsive polymer. 2. The method of claim 1, wherein the magnetic field does not induce magnetophoresis in a non-aggregated stimuli-responsive magnetic nanoparticle in the liquid. 3. The method of claim 1 further comprising a step of concentrating the aggregate in the liquid. 4. The method of claim 1, wherein the first stimuli-responsive polymer and the second stimuli-responsive polymer are responsive to stimuli independently selected from the group consisting of temperature, pH, light, photo-irradiation, exposure to an electric field, ionic strength, and combinations thereof. 5. The method of claim 1, wherein the stimuli-responsive capture conjugate further comprises a detector complex comprising a detector moiety and a second capture moiety, wherein the second capture moiety is capable of binding to the target. 6. The method of claim 1, wherein the target is an indicator for a disease or condition. 7. The method of claim 1, wherein the target is selected from the group consisting of an antibody, an antigen, a cell, a nucleic acid, an enzyme, a substrate for an enzyme, a protein, a lipid, a carbohydrate, or other biomarker. 8. The method of claim 1 further comprising a biological sample selected from the group consisting of blood, mucus, urine, tissue, sputum, saliva, feces, a nasal swab, and nasopharyngeal washes. 9. The method of claim 1, wherein the magnetic core is a metal oxide. 10. The method of claim 1 further comprising a second stimuli-responsive capture conjugate comprising a third stimuli-responsive polymer attached to a third capture moiety, wherein the third capture moiety is capable of capturing a second target, wherein the second target is different than the target. 11. A stimuli-responsive reagent, comprising: (a) a stimuli-responsive magnetic nanoparticle comprising a first stimuli-responsive polymer attached to a magnetic core; and(b) a stimuli-responsive capture conjugate comprising a second stimuli-responsive polymer attached to a first capture moiety, wherein the first capture moiety is capable of binding to a target. 12. The stimuli-responsive reagent of claim 11, wherein the first stimuli-responsive polymer and the second stimuli-responsive polymer are responsive to stimuli independently selected from the group consisting of temperature, pH, light, photo-irradiation, exposure to an electric field, ionic strength, and combinations thereof. 13. The stimuli-responsive reagent of claim 11, wherein the stimuli-responsive reagent further comprises a detector complex comprising a detector moiety and a second capture moiety, wherein the second capture moiety is capable of binding to the target. 14. The stimuli-responsive reagent of claim 11, wherein the target is an indicator for a disease or condition. 15. The stimuli-responsive reagent of claim 11, wherein the target is selected from the group consisting of an antibody, an antigen, a cell, a nucleic acid, an enzyme, a substrate for an enzyme, a protein, a lipid, a carbohydrate, or other biomarker. 16. The stimuli-responsive reagent of claim 11, wherein the magnetic core is a metal oxide. 17. The stimuli-responsive reagent of claim 11 further comprising a second stimuli-responsive capture conjugate comprising a third stimuli-responsive polymer attached to a third capture moiety, wherein the third capture moiety is capable of capturing a second target, wherein the second target is different than the target. 18. A method of capturing a target in a liquid, comprising: (a) contacting a liquid to be tested for the presence of a target with a stimuli-responsive reagent for a pre-determined period of time sufficient to effect binding of the target, if present, to the stimuli-responsive capture conjugate, comprising: (i) a stimuli-responsive magnetic nanoparticle comprising a first stimuli-responsive polymer attached to a magnetic core; and(ii) a stimuli-responsive capture conjugate comprising a second stimuli-responsive polymer attached to a capture moiety, wherein the capture moiety is capable of binding to a target;(b) applying an effective stimulus to provide an aggregate in the liquid formed through associative interaction between the first stimuli-responsive polymer and the second stimuli-responsive polymer, wherein the aggregate comprises: (i) a stimuli-responsive magnetic nanoparticle comprising a first stimuli-responsive polymer attached to a magnetic core; and(ii) a stimuli-responsive capture conjugate comprising a second stimuli-responsive polymer attached to a capture moiety, wherein the capture moiety is bound to the target when the target is present in the liquid,(c) subjecting the aggregate to a magnetic field to magnetophorese the aggregate to a site within the liquid to provide a magnetophoresed aggregate in the liquid; and(d) analyzing the magnetophoresed aggregate to determine if the target is present. 19. The method of claim 18 further comprising binding a detector complex to the target, said detector complex comprising a second capture moiety capable of binding to the target and a reporting moiety. 20. The method of claim 19, wherein the step of analyzing the magnetophoresed aggregate comprises photoluminescence analysis.
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