Method of analysis for determining a specific protein in blood samples using fluorescence spectrometry
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
G01N-033/00
G01N-033/58
출원번호
US-0767196
(2007-06-22)
등록번호
US-9040305
(2015-05-26)
발명자
/ 주소
Goix, Philippe J.
Puskas, Robert
Todd, John
Livingston, Richard A.
Held, Douglas
출원인 / 주소
Singulex, Inc.
대리인 / 주소
McDonnell Boehnen Hulbert & Berghoff LLP
인용정보
피인용 횟수 :
1인용 특허 :
128
초록▼
The invention encompasses analyzers and analyzer systems that include a single particle analyzer, methods of using the analyzers and analyzers systems to analyze samples, either for single particles, e.g., protein molecules, or for multiple particles (multiplexing), methods of doing business based o
The invention encompasses analyzers and analyzer systems that include a single particle analyzer, methods of using the analyzers and analyzers systems to analyze samples, either for single particles, e.g., protein molecules, or for multiple particles (multiplexing), methods of doing business based on the use of the analyzers or analyzer systems of the system, and electronic media for storing parameters useful in the analyzers and analyzer systems of the invention.
대표청구항▼
1. A method of analysis for determining the presence, absence or amount of a specific protein in a blood sample, comprising the steps of: (a) providing a processed sample, wherein the processed sample has been prepared by (i) extracting serum or plasma from the blood sample;(ii) contacting the serum
1. A method of analysis for determining the presence, absence or amount of a specific protein in a blood sample, comprising the steps of: (a) providing a processed sample, wherein the processed sample has been prepared by (i) extracting serum or plasma from the blood sample;(ii) contacting the serum or plasma with a selective capture agent specific for the protein, wherein the capture agent is immobilized or becomes immobilized on a surface;(iii) adding a fluorescently-labeled specific binding partner for the protein, wherein the binding partner and the protein associate to form a protein-binding partner complex;(iv) removing labeled binding partner that has not associated with the protein;(v) releasing the protein-binding partner complex into solution or dissociating the labeled binding partner from the protein-binding partner complex so that the dissociated labeled binding partner moves into solution;(b) sampling a portion of the processed sample, wherein the sampling is performed by a sampling system that comprises a pressure source to move the portion of the processed sample into a detection channel of a single molecule analyzer, wherein the detection channel is substantially transparent to at least some wavelengths of light and provides a channel through which the portion of the processed sample may flow, and the analyzer comprises a laser for providing excitation light, wherein the light is within the wavelengths to which the detection channel is substantially transparent; and(c) passing the portion of the processed sample through the detection channel of the analyzer using positive or negative pressure;(d) focusing the excitation light on a portion of the detection channel, so that the light excites the labeled specific binding partner if present in the portion of the detection channel so that the labeled specific binding partner produces emitted light;(e) passing the emitted light through an aperture so that the focusing of excitation light and passing of emitted light through the aperture define a molecule detection volume within the portion of the detection channel of between about 0.1 pL and about 25 pL;(f) detecting, in discreet time bins, the emitted light that has passed through the aperture with a detector, wherein the detector transforms the light into an electronic signal; and(g) determining the presence, absence or amount of the specific protein in a blood sample by determining a threshold photon bin value based upon a distribution of background noise signal and analyzing the electronic signal from a plurality of the bins with a data analysis system operably connected to the detector that compares the signal for each bin to the threshold value to determine whether or not the labeled specific binding partner is present in the molecule detection volume. 2. The method of claim 1 wherein the protein is present in the sample at a concentration of about 100 femtomolar to about 1500 femtomolar. 3. The method of claim 1 wherein the protein is present in the sample at a concentration of 0.5-10 pg/ml. 4. The method of claim 1 wherein light emitted from the molecule detection volume is associated with a single labeled specific binding partner that has been excited by the excitation light. 5. The method of claim 1 wherein the labeled specific binding partner is associated with a particle, and the light emitted from the detection volume is associated with a single particle that has been excited by excitation light. 6. The method of claim 1 wherein the labeled specific binding partner has low photobleaching. 7. The method of claim 1, wherein the threshold value assumes a Poission distribution of the background noise signal. 8. The method of claim 1, wherein the threshold level is a set a fixed number of standard deviations above the background signal. 9. The method of claim 8, wherein the data analysis system determines a signal above the threshold level as a single labeled specific binding partner so that each bin is analyzed as a “yes” or “no” for the presence of the labeled specific binding partner. 10. The method of claim 1, wherein the bin time is longer than the time the labeled specific binding partner passes through the detection volume. 11. A method of determining the presence, absence or amount of an analyte comprising a specific binding substance in a sample, the method comprising: (a) contacting the sample with a capture agent specific for the analyte and a fluorescently-labeled specific binding partner for the analyte to form a complex comprising the capture agent, the analyte and the labeled binding partner, wherein the capture agent is immobilized or becomes immobilized on a surface;(b) removing labeled binding partner that has not associated with the analyte;(c) releasing the analyte or the labeled binding partner from the complex so that the dissociated labeled binding partner moves into solution;(d) passing the labeled binding partner through a detection channel of an analyzer, wherein the detection channel comprises a portion that is substantially transparent to at least some wavelengths of light and provides a channel through which the labeled binding partner may flow, and the analyzer comprises a laser for providing excitation light, wherein the light is within the wavelengths to which the portion of the detection channel is substantially transparent;(d) focusing the excitation light within the portion of the detection channel, so that the light excites the labeled specific binding partner if present in the portion of the detection channel so that the labeled specific binding partner produces emitted light;(e) passing the emitted light through an aperture so that the focusing of excitation light and passing of emitted light through the aperture define a detection volume within the portion of the detection channel of between about 0.1 pL and about 25 pL;(f) detecting, in discreet time bins, the emitted light that has passed through the aperture with a detector, wherein the detector transforms the light into an electronic signal; and(g) determining the presence, absence or amount of the analyte in the sample by determining a threshold photon bin value based upon a distribution of background noise signal and analyzing the electronic signal from a plurality of the bins with a data analysis system operably connected to the detector that compares the signal for each bin to the threshold value to determine whether or not the labeled specific binding partner is present in the detection volume. 12. The method of claim 11, wherein the threshold value assumes a Poission distribution of the background noise signal. 13. The method of claim 11, wherein the threshold level is a set a fixed number of standard deviations above the background signal. 14. The method of claim 11, wherein the data analysis system determines a signal above the threshold level as single labeled specific binding partner so that each bin is analyzed as a “yes” or “no” for the presence of the labeled specific binding partner. 15. The method of claim 11, wherein the determining the presence, absence or amount of the analyte in the sample further comprises determining the number of bins having a photon bin count greater than the threshold level for a sample comprising a standard, and comparing the number of bins having a photon bin count greater to a threshold level for the sample comprising the analyte to the number of bins having a photon bin count greater than a threshold level for the sample comprising the standard. 16. The method of claim 11, wherein the bin time is longer than the time the labeled specific binding partner passes through the detection volume.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (128)
Heller Michael J. (Encinitas CA) Tu Eugene (San Diego CA), Active programmable electronic devices for molecular biological analysis and diagnostics.
Chang, Richard K.; Pan, Young-Le; Pinnick, Ronald G.; Hill, Steven C., Aerosol fluorescence spectrum analyzer for rapid measurement of single airborne particles.
Baer Thomas M. (Mountain View CA) Dietz Louis J. (Mountain View CA) Dubrow Robert S. (San Carlos CA) Hayter Paul G. (Los Altos CA) Hodges Michael (Palo Alto CA) Manian Bala S. (Los Altos Hills CA) Sh, Apparatus and method for volumetric capillary cytometry.
Groman Ernest V. (Brookline MA) Josephson Lee (Arlington MA), Biologically degradable superparamagnetic particles for use as nuclear magnetic resonance imaging agents.
Mathies Richard A. (Contra Costa County CA) Huang Xiaohua C. (Santa Clara County CA) Quesada Mark A. (San Francisco County CA), Capillary array confocal fluorescence scanner and method.
Chow Calvin Y. H. ; Parce John Wallace ; McReynolds Richard J. ; Kennedy Colin B. ; Bousse Luc J., Controller/detector interfaces for microfluidic systems.
Hoffman Michael A. (Carmel CA), Fluorescent-particle analyzer with timing alignment for analog pulse subtraction of fluorescent pulses arising from diff.
Wada, H. Garrett; Kopf-Sill, Anne R.; Alajoki, Marja Liisa; Parce, J. Wallace; Wang, Benjamin N.; Chow, Andrea W.; Dubrow, Robert S., Focusing of microparticles in microfluidic systems.
Swedberg Sally A. (Los Altos CA) Kaltenbach Patrick (Bischweier DEX) Witt Klaus E. (Keltern DEX) Bek Fritz (Waldbronn DEX) Mittelstadt Laurie S. (Belmont CA), Fully integrated miniaturized planar liquid sample handling and analysis device.
Dollinger Gavin D. (San Francisco CA) Cunico Robert L. (Hercules CA) Kunitani Michael G. (San Rafael CA), HPLC light scattering detector for biopolymers.
Mathies Richard A. (Contra Costa County CA) Peck Konan (Contra Costa County CA) Stryer Lubert (Santa Clara County CA), High sensitivity fluorescent single particle and single molecule detection apparatus and method.
Ambrose W. Patrick ; Grace W. Kevin ; Goodwin Peter M. ; Jett James H. ; Orden Alan Van ; Keller Richard A., High throughput analysis of samples in flowing liquid.
Gourley Paul L. (12508 Loyola ; NE. Albuquerque NM 87112) Gourley Mark F. (7509 Spring Lake Dr. ; Apt. B1 Bethesda MD 20817), Laser apparatus and method for microscopic and spectroscopic analysis and processing of biological cells.
Witt Klaus E. (Keltern DEX) Kaltenbach Patrick (Bischweier DEX) Bek Fritz (Waldbronn DEX) Swedberg Sally A. (Los Altos CA) Mittelstadt Laurie S. (Belmont CA), Low voltage miniaturized column analytical apparatus and method.
Miltenyi Stefan (Moitzfeld 60a ; D-5060 Bergisch Gladbach 1 Cologne DEX) Radbruch Andreas (Bonn DEX) Weichel Walter (Cologne DEX) Muller Werner (Cologne DEX) Gottlinger Christoph (Cologne DEX) Meyer , Metal matrices for use in high gradient magnetic separation of biological materials and method for coating the same.
Begg Geoffrey Stephen (Heidelberg AUX) Simpson Richard John (Richmond AUX) Burgess Antony Wilks (Camberwell AUX), Method allowing sequential chemical reactions.
Purvis ; Jr. Norman B. (1010 Lawnview Ct. Franklin TN 37064) Giorgio Todd D. (3608 Westbrook Dr. Nashville TN 37205), Method and apparatus for determining absolute particle size, surface area and volume normalized fluorescence using forwa.
Cherukuri Satyam C. (Cranbury NJ) Demers Robert R. (Cranbury NJ) Fan Zhong H. (Middlesex NJ) Levine Aaron W. (Lawrenceville NJ) McBride Sterling E. (Lawrence Township ; Mercer County NJ) Zanzucchi Pe, Method and system for inhibiting cross-contamination in fluids of combinatorial chemistry device.
Eigen Manfred,DEX ; Winkler Thorsten,DEX ; Stephan Jens,DEX ; Schwille Petra,DEX ; Koltermann Andre,DEX ; Kettling Ulrich,DEX ; Dorre Klaus,DEX ; Bieschke Jan,DEX, Method for detecting reactions by means of coincidence analysis.
Seidel Claus,DEX ; Gunther Rolf,DEX ; Lupke Stefan,DEX, Method for differentiating or detecting particles in a sample by identifying signal segments of time-resolved, optical raw signals from the sample on the basis of single photon detection.
Goix, Philippe J.; Puskas, Robert; Todd, John; Livingston, Richard A.; Held, Douglas, Method for highly sensitive detection of single protein molecules labeled with fluorescent moieties.
Robotti, Karla M.; Yin, Hongfeng, Methods and using chemico-mechanical microvalve devices for the selective separation of components from multi-component fluid samples.
Fauver, Mark E.; Seibel, Eric J.; Brown, Chris M.; Reinhall, Per G.; Smithwick, Quinn Y. J., Micro-fabricated optical waveguide for use in scanning fiber displays and scanned fiber image acquisition.
Richard A. Mathies ; Pankaj Singhal ; Jin Xie ; Alexander N. Glazer, Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels.
Kaltenbach Patrick,DEX ; Swedberg Sally A. ; Witt Klaus E.,DEX ; Bek Fritz,DEX ; Mittelstadt Laurie S., Miniaturized planar columns for use in a liquid phase separation apparatus.
Kaltenbach Patrick (Bischweier DEX) Swedberg Sally A. (Los Altos CA) Witt Klaus E. (Keltern DEX) Bek Fritz (Waldbronn DEX) Mittelstadt Laurie S. (Belmont CA), Miniaturized planar columns in novel support media for liquid phase analysis.
Hollis Mark A. (Concord MA) Ehrlich Daniel J. (Lexington MA) Murphy R. Allen (Boxboro MA) Kosicki Bernard B. (Acton MA) Rathman Dennis D. (Ashland MA) Chen Chang-Lee (Sudbury MA) Mathews Richard H. (, Optical and electrical methods and apparatus for molecule detection.
Goodwin Peter M. ; Jett James H. ; Keller Richard A. ; Van Orden Alan K. ; Machara Nicholas P., Single molecule identification using selected fluorescence characteristics.
Galambos, Paul C.; Okandan, Murat; Montague, Stephen; Smith, James H.; Paul, Phillip H.; Krygowski, Thomas W.; Allen, James J.; Nichols, Christopher A.; Jakubczak, II, Jerome F., Surface-micromachined microfluidic devices.
Doth Margit,DEX ; Petry Christoph,DEX, Synthetic calibrators for use in immunoassays, comprising the analytes or partial sequences thereof which are conjugated to inert carrier molecules.
Myers Stephen A. (25 Nimitz Pl. Old Greenwich CT 06870), System and method for determining changes in fluorescence of stained nucleic acid in electrophoretically separated bands.
Muller Ralph,DEX ; Sauer Markus,DEX ; Zander Christoph,DEX, System for distinguishing fluorescent molecule groups by time resolved fluorescence measurement.
Goix, Philippe; Puskas, Robert; Todd, John; Livingston, Richard; Held, Douglas; Wu, Allan H. B., Highly sensitive system and method for analysis of troponin.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.