System and method for magnetically concentrating and detecting biomarkers
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12M-001/00
G01N-031/00
G01N-031/22
G01N-009/00
출원번호
US-0815217
(2010-06-14)
등록번호
US-8426214
(2013-04-23)
발명자
/ 주소
Stayton, Patrick S.
Nash, Michael
Lai, Jriuan
출원인 / 주소
University of Washington
대리인 / 주소
Christensen O'Connor Johnson Kindness PLLC
인용정보
피인용 횟수 :
8인용 특허 :
46
초록▼
System and method for capturing, concentrating, and detecting a diagnostic target in a liquid, comprising applying a magnetic field to a mixture comprising a co-aggregate in the liquid to provide a collected co-aggregate in the liquid, wherein the co-aggregate comprises a magnetic particle having a
System and method for capturing, concentrating, and detecting a diagnostic target in a liquid, comprising applying a magnetic field to a mixture comprising a co-aggregate in the liquid to provide a collected co-aggregate in the liquid, wherein the co-aggregate comprises a magnetic particle having a stimuli-responsive polymer attached thereto and a non-magnetic particle having a stimuli-responsive polymer and a diagnostic target attached thereto.
대표청구항▼
1. A method for concentrating a non-magnetic particle in a liquid, comprising: (a) applying a magnetic field to a mixture comprising a co-aggregate in a liquid to provide a collected co-aggregate in the liquid, wherein the co-aggregate comprises: (i) a magnetic particle having a stimuli-responsive p
1. A method for concentrating a non-magnetic particle in a liquid, comprising: (a) applying a magnetic field to a mixture comprising a co-aggregate in a liquid to provide a collected co-aggregate in the liquid, wherein the co-aggregate comprises: (i) a magnetic particle having a stimuli-responsive polymer attached thereto; and(ii) a non-magnetic particle having a stimuli-responsive polymer attached thereto;wherein the non-magnetic particle further comprises a first binding partner of a binding pair, wherein the liquid comprises a second binding partner of the binding pair, and wherein the co-aggregate comprises a non-magnetic particle having the first and second binding partners attached thereto;(b) removing at least a portion of the liquid from the collected co-aggregate in the liquid;(c) re-dissolving the co-aggregate to provide a mixture comprising the magnetic particle having the stimuli-responsive polymer attached thereto and the non-magnetic particle having the stimuli-responsive polymer and first and second binding partners attached thereto; and(d) contacting the mixture comprising the magnetic particle having the stimuli-responsive polymer attached thereto and the non-magnetic particle having the stimuli-responsive polymer and first and second binding partners attached thereto with a solid phase effective to immobilize the non-magnetic particle having the stimuli-responsive polymer and first and second binding partners attached thereto. 2. The method of claim 1, wherein the co-aggregate further comprises free stimuli-responsive polymer. 3. A method for capturing a diagnostic target in a liquid medium, comprising: (a) contacting a liquid medium to be tested for the presence of a diagnostic target with a plurality of stimuli-responsive particles, wherein the plurality of stimuli-responsive particles comprises (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto, and(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer and a capture moiety attached thereto, wherein the capture moiety has an affinity to the diagnostic target,wherein the plurality of stimuli-responsive particles is contacted with the liquid medium for a pre-determined period of time sufficient to effect binding of the diagnostic target, if present, to a portion of the plurality of the non-magnetic particles; andwherein the liquid medium to be tested comprises a first and a second diagnostic target and the plurality of non-magnetic particles comprises(i) a plurality of first non-magnetic particles, each having a stimuli-responsive polymer and a first capture moiety attached thereto, wherein the first capture moiety has an affinity to the first diagnostic target, and(ii) a plurality of second non-magnetic particles, each having a stimuli-responsive polymer and a second capture moiety attached thereto, wherein the second capture moiety has an affinity to the second diagnostic target;(b) applying an external stimulus to provide a co-aggregate in the liquid medium, wherein the co-aggregate comprises (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto, and(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer thereto, wherein at least a portion of the plurality of non-magnetic particles have the diagnostic target attached thereto when the diagnostic target is present in the liquid medium, andwherein the co-aggregate is formed by the association of the stimuli-responsive polymer attached to the magnetic particle to the stimuli-responsive polymer attached to the non-magnetic particle;(c) subjecting the co-aggregate to a magnetic field to magnetophorese the co-aggregate to a site within the liquid medium to provide a magnetophoresced co-aggregate in the liquid medium;(d) removing at least portion of the liquid medium from the liquid medium comprising the magnetophoresced co-aggregate to provide the co-aggregate and optionally residual liquid medium;(e) removing the stimulus and the magnetic field to provide a mixture comprising (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto,(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer thereto, wherein at least a portion of the plurality of non-magnetic particles have the diagnostic target attached thereto when the diagnostic target is present in the liquid medium, and(iii) optionally residual liquid medium; and(f) contacting the mixture with a solid phase to immobilize at least a portion of the first plurality of non-magnetic particles having the first diagnostic target attached thereto and to immobilize at least a portion of the second plurality of non-magnetic particles having the second diagnostic target attached thereto. 4. The method of claim 3 further comprising contacting the mixture with a solid phase to immobilize at least a portion of the plurality of non-magnetic particles having the diagnostic target attached thereto. 5. The method of claim 3, wherein the diagnostic target is an antigen and the capture moiety is an antibody. 6. The method of claim 3, wherein the diagnostic target is a biomarker for a disease or disorder. 7. The method of claim 3, wherein the non-magnetic particles having the first diagnostic target attached thereto are immobilized on the solid phase at a first position and the non-magnetic particles having the second diagnostic target attached thereto are immobilized on the solid phase at a second position, wherein the first and second positions are not the same. 8. A method for capturing a diagnostic target in a liquid medium, comprising: (a) contacting a liquid medium to be tested for the presence of a diagnostic target with a first binding partner having an affinity to the diagnostic target and plurality of stimuli-responsive particles, wherein the plurality of stimuli-responsive particles comprises (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto, and(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer and a second binding partner attached thereto, wherein the second binding partner has an affinity to the first binding partner,wherein the plurality of stimuli-responsive particles is contacted with the liquid medium for a pre-determined period of time sufficient to effect binding of the diagnostic target, if present, to a portion of the plurality of non-magnetic particles through the association of the first and second binding partners;(b) applying an external stimulus to provide a co-aggregate in the liquid medium, wherein the co-aggregate comprises (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto, and(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer thereto, wherein at least a portion of the plurality of non-magnetic particles have the diagnostic target attached thereto when the diagnostic target is present in the liquid medium, andwherein the co-aggregate is formed by the association of the stimuli-responsive polymer attached to the magnetic particle to the stimuli-responsive polymer attached to the non-magnetic particle;(c) subjecting the co-aggregate to a magnetic field to magnetophorese the co-aggregate to a site within the liquid medium to provide a magnetophoresced co-aggregate in the liquid medium;(d) removing at least portion of the liquid medium from the liquid medium comprising the magnetophoresced co-aggregate to provide the co-aggregate and optionally residual liquid medium;(e) removing the stimulus and the magnetic field to provide a mixture comprising (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto,(ii) a plurality of non-magnetic particles, each having a stimuli-responsive polymer thereto, wherein at least a portion of the plurality of non-magnetic particles have the diagnostic target attached thereto when the diagnostic target is present in the liquid medium, and(iii) optionally residual liquid medium; and(f) contacting the mixture with a solid phase to immobilize at least a portion of the plurality of non-magnetic particles have the diagnostic target attached thereto. 9. The method of claim 8, wherein the liquid medium to be tested for a diagnostic target is to be tested for first and second diagnostic targets, wherein contacting the liquid medium comprises contacting with a first binding partner having an affinity to the first diagnostic target, a third binding partner having an affinity to the second diagnostic target, and plurality of stimuli-responsive particles, wherein the plurality of stimuli-responsive particles comprises (i) a plurality of magnetic particles, each having a stimuli-responsive polymer attached thereto,(ii) a plurality of first non-magnetic particles, each having a stimuli-responsive polymer and a second binding partner attached thereto, wherein the second binding partner has an affinity to the first binding partner, and(iii) a plurality of second non-magnetic particles, each having a stimuli-responsive polymer and a fourth binding partner attached thereto, wherein the fourth binding partner has an affinity to the third binding partner,wherein the plurality of stimuli-responsive particles is contacted with the liquid medium for a pre-determined period of time sufficient to effect binding of the first and second diagnostic targets, if present, to a portion of the pluralities of the first and second non-magnetic particles through the association of the first and second, and third and fourth binding partners, respectively. 10. The method of claim 9 further comprising contacting the mixture with a solid phase to immobilize at least a portion of the plurality of first non-magnetic particles having the first diagnostic target attached thereto and to immobilize at least a portion of the plurality of second non-magnetic particles having the second diagnostic target attached thereto. 11. The method of claim 10, wherein the non-magnetic particles having the first diagnostic target attached thereto are immobilized on the solid phase at a first position and the non-magnetic particles having the second diagnostic target attached thereto are immobilized on the solid phase at a second position, wherein the first and second positions are not the same.
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