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WO1994017409B1 - Quantifying trace analytes by affinity capillary electrophoresis - Google Patents

Quantifying trace analytes by affinity capillary electrophoresis

Info

Publication number
WO1994017409B1
WO1994017409B1 PCT/US1994/000426 US9400426W WO9417409B1 WO 1994017409 B1 WO1994017409 B1 WO 1994017409B1 US 9400426 W US9400426 W US 9400426W WO 9417409 B1 WO9417409 B1 WO 9417409B1
Authority
WO
WIPO (PCT)
Prior art keywords
analyte
labelled
fragment
fab
agent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1994/000426
Other languages
French (fr)
Other versions
WO1994017409A1 (en
Filing date
Publication date
Priority claimed from US08/006,922 external-priority patent/US5348633A/en
Application filed filed Critical
Priority to AU61625/94A priority Critical patent/AU6162594A/en
Priority to JP51708594A priority patent/JP3530914B2/en
Priority to EP94908588A priority patent/EP0680608A1/en
Publication of WO1994017409A1 publication Critical patent/WO1994017409A1/en
Publication of WO1994017409B1 publication Critical patent/WO1994017409B1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Abstract

A method for quantitative detection of trace amounts of an analyte in a sample is disclosed. The method in the preferred embodiment includes providing an Fab' fragment of an immunoglobulin labelled at a reactive sulfhydryl group with a fluorescent dye; combining the labelled Fab' fragment with a sample that may contain the analyte; concentrating the elements of the resulting mixture in an electric field; separating the labelled analyte/agent complex formed from any unreacted labelled agent using capillary electrophoretic methods; and detecting the fluorescent signal of the labelled analyte/agent complex. The invention also is directed to a method of producing a labelled Fab' fragment that includes cleaving an immunoglobulin molecule to obtain one F(ab')2 fragment; reducing the disulfide-bonds of the F(ab')2 fragment to obtain two Fab' fragments each having at least one free, reactive sulfhydryl group; and mixing an Fab' fragment having at least one free sulfhydryl group with a fluorescent dye reactive with the free sulfhydryl to form a labelled Fab' fragment. Preferably, prior to the final mixing step, intrastrand disulfide bonds are formed by oxidation within each Fab' fragment, thereby producing individual Fab' fragments each having a single reactive sulfhydryl group. The method of quantitative detection also more broadly includes using any biospecific agent to form a complex with the target analyte.

Claims

/17409 P
AMENDED CLAIMS
I received by the International Bureau on 5 August 1994 (05.08.94); original claim 21 amended; other claims unchanged (1 page)]
20. The method of claim 18, wherein said detectable reporter group is not sensitive to pH change in the pH range of operation.
21. A method for quantitatively detecting trace amounts of an analyte in a sample comprising the steps of: mixing together a sample which may contain said analyte and a biospecific agent that can bind to said analyte to form an analyte-agent complex, said analyte-agent complex containing a detectable reporter group; concentrating elements of said mixture in an electric field by a capillary electrophoretic concentration technique; separating elements of said mixture by a capillary electrophoretic separation technique; and quantitatively detecting said reporter group on said analyte-agent complex as an indication of the amount of said analyte in said sample.
22. The method of claim 21, wherein said biospecific agent is labelled with said detectable reporter group.
23. The method of claim 21 further comprising in said mixing step, mixing a known amount of labelled said analyte, said labelled analyte containing said detectable reporter group.
24. The method of claim 21, wherein said biospecific agent is selected from the group consisting of antibody fragment, binding protein, and ligand.
25. The method of claim 21, wherein said reporter group is selected from the group consisting of fluorophore, UV or visible chromophore, radioisotope, spin label, electrochemical reporter, chemiluminescent reporter, and enzyme.
PCT/US1994/000426 1993-01-22 1994-01-12 Quantifying trace analytes by affinity capillary electrophoresis Ceased WO1994017409A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU61625/94A AU6162594A (en) 1993-01-22 1994-01-12 Quantifying trace analytes by affinity capillary electrophoresis
JP51708594A JP3530914B2 (en) 1993-01-22 1994-01-12 Determination of trace analytes by affinity capillary electrophoresis.
EP94908588A EP0680608A1 (en) 1993-01-22 1994-01-12 Quantifying trace analytes by affinity capillary electrophoresis

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/006,922 US5348633A (en) 1993-01-22 1993-01-22 Method for quantitating trace amounts of an analyte in a sample by affinity capillary electrophoresis
US006,922 1993-01-22

Publications (2)

Publication Number Publication Date
WO1994017409A1 WO1994017409A1 (en) 1994-08-04
WO1994017409B1 true WO1994017409B1 (en) 1994-09-15

Family

ID=21723287

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1994/000426 Ceased WO1994017409A1 (en) 1993-01-22 1994-01-12 Quantifying trace analytes by affinity capillary electrophoresis

Country Status (5)

Country Link
US (1) US5348633A (en)
EP (1) EP0680608A1 (en)
JP (1) JP3530914B2 (en)
AU (1) AU6162594A (en)
WO (1) WO1994017409A1 (en)

Families Citing this family (49)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5468359A (en) * 1988-11-14 1995-11-21 Anthony R. Torres Method of determining presence of an analyte by isoelectric focusing
US5571680A (en) * 1994-01-21 1996-11-05 Beckman Instruments, Inc. Homogeneous immunoassays and enzyme based assays of analytes using capillary electrophoresis
US5536382A (en) * 1994-05-23 1996-07-16 Advanced Molecular Systems, Inc. Capillary electrophoresis assay method useful for the determination of constituents of a clinical sample
US5431793A (en) * 1994-07-29 1995-07-11 Beckman Instruments, Inc. Quantitative analysis of glycosylated hemoglobin by immunocappillary electrophoresis
EP0778777A4 (en) * 1994-08-22 1999-01-27 Univ Pennsylvania A method of sequencing proteins by epitope ordering and protein restriction mapping
WO1996010170A1 (en) * 1994-09-29 1996-04-04 The Board Of Trustees Of The Leland Stanford Junior University Capillary-based separation methods for identifying bioactive analytes in a mixture
GB9421468D0 (en) * 1994-10-18 1994-12-07 Surface Active Ltd Biosensor
DE69605712T2 (en) * 1995-03-31 2000-04-27 Laboratory Of Molecular Biophotonics, Hamakita Application of isoelectric point markers in isoelectric focusing with fluorescence detection
US5630924A (en) * 1995-04-20 1997-05-20 Perseptive Biosystems, Inc. Compositions, methods and apparatus for ultrafast electroseparation analysis
US5783397A (en) * 1995-12-11 1998-07-21 Northeastern University Screening natural samples for new therapeutic compounds using capillary electrophoresis
US5824478A (en) * 1996-04-30 1998-10-20 Vysis, Inc. Diagnostic methods and probes
US6299747B1 (en) 1997-12-17 2001-10-09 Cetek Corporation Capillary electrophoretic methods to detect new biologically active compounds in complex biological material
US6406604B1 (en) 1999-11-08 2002-06-18 Norberto A. Guzman Multi-dimensional electrophoresis apparatus
US7329388B2 (en) 1999-11-08 2008-02-12 Princeton Biochemicals, Inc. Electrophoresis apparatus having staggered passage configuration
AU3102801A (en) * 2000-01-19 2001-07-31 Exact Sciences Corporation Methods of and apparatus for separating and detecting nucleic acid
JP3461804B2 (en) * 2000-02-10 2003-10-27 浜松ホトニクス株式会社 Method for producing Fab 'antibody with homogenized isoelectric point and Fab' antibody with homogenized fluorescent label
JP3439195B2 (en) * 2000-02-17 2003-08-25 浜松ホトニクス株式会社 Antigen quantitative detection method
DE10127045C2 (en) * 2001-06-02 2003-10-30 November Ag Molekulare Medizin Method for the detection of a substance and microtiter plate
WO2002101389A1 (en) * 2001-06-12 2002-12-19 Wako Pure Chemical Industries, Ltd. Method of assaying hyaluronic acid
US7223538B2 (en) * 2001-12-14 2007-05-29 Ge Healthcare Bio-Sciences Ab Post-synthesis labeling of nucleic acids, assays using nucleic acids that are labeled post-synthetically, single nucleotide polymorphism detection, and associated compounds and microarrays
US20050211555A1 (en) * 2002-02-14 2005-09-29 Solus Biosystems, Inc. Method for multiple sample screening using IR spectroscopy
WO2004048929A2 (en) * 2002-11-22 2004-06-10 Solus Biosystems, Inc. High throughput screening with parallel vibrational spectroscopy
US20040260414A1 (en) * 2003-06-20 2004-12-23 Groton Biosystems, Llc Method and apparatus for operating an automated biomolecular preparation system
US7601545B2 (en) * 2003-06-20 2009-10-13 Groton Biosystems, Llc Automated macromolecule sample preparation system
US8007724B2 (en) * 2003-11-07 2011-08-30 Princeton Biochemicals, Inc. Electrophoresis apparatus having at least one auxiliary buffer passage
WO2005047882A2 (en) * 2003-11-07 2005-05-26 Princeton Biochemicals, Inc. Multi-dimensional electrophoresis apparatus
US20050130319A1 (en) * 2003-12-15 2005-06-16 Xerox Corporation Molecular binding event detection using separation channels
EP1697732A1 (en) * 2003-12-23 2006-09-06 Caliper Life Sciences, Inc. Analyte injection system
US7407816B2 (en) * 2004-05-07 2008-08-05 Gentius, Inc Isoelectric particles and uses thereof
US20060292649A1 (en) * 2004-07-19 2006-12-28 Cell Biosciences Inc. Methods and apparatus for reference lab diagnostics
EP1776581B1 (en) * 2004-07-19 2015-05-06 ProteinSimple Method for protein detection
US7846676B2 (en) * 2004-07-19 2010-12-07 Cell Biosciences, Inc. Methods and devices for analyte detection
US20060292558A1 (en) * 2004-07-19 2006-12-28 Cell Biosciences Inc. Methods and apparatus for protein assay diagnostics
US7935479B2 (en) 2004-07-19 2011-05-03 Cell Biosciences, Inc. Methods and devices for analyte detection
WO2006078366A2 (en) * 2005-01-18 2006-07-27 Solus Biosystems, Inc. Multiple sample screening using ir spectroscopy with capillary isoelectric focusing
JP2008536128A (en) * 2005-04-09 2008-09-04 セル バイオサイエンシズ，インコーポレイテッド Automated microvolume assay system
US20060286378A1 (en) * 2005-05-23 2006-12-21 Shivkumar Chiruvolu Nanostructured composite particles and corresponding processes
CA2620403A1 (en) * 2005-09-02 2007-03-08 Wako Pure Chemical Industries, Ltd. Complex formation method and separation method
JP4724816B2 (en) * 2005-09-06 2011-07-13 シャープ株式会社 Protein measurement method
WO2007035864A2 (en) * 2005-09-20 2007-03-29 Cell Biosciences, Inc. Electrophoresis standards, methods and kits
US9081006B2 (en) * 2007-02-07 2015-07-14 Lyzer Diagnostics, Inc. Rapid homogeneous immunoassay using electrophoresis
US20090023156A1 (en) * 2007-07-20 2009-01-22 Voss Karl O Methods and reagents for quantifying analytes
US10107782B2 (en) 2008-01-25 2018-10-23 ProteinSimple Method to perform limited two dimensional separation of proteins and other biologicals
EP2283347A1 (en) * 2008-05-27 2011-02-16 Koninklijke Philips Electronics N.V. Biochip for fractionating and detecting analytes
GB201010405D0 (en) 2010-06-21 2010-08-04 Binding Site Group The Ltd Assay
US9804079B2 (en) 2012-04-19 2017-10-31 ProteinSimple Dual wavelength isoelectric focusing for determining drug load in antibody drug conjugates
US9766206B2 (en) 2013-09-27 2017-09-19 ProteinSimple Apparatus, systems, and methods for capillary electrophoresis
AU2024272220A1 (en) * 2023-05-12 2025-10-16 Novo Nordisk A/S Long-acting growth hormone receptor antagonist and use thereof
WO2025157168A1 (en) * 2024-01-23 2025-07-31 Wuxi Biologics (Shanghai) Co., Ltd. Imaged capillary isoelectric focusing for detecting mispairing byproduct in asymmetric bispecific antibodies

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4725343A (en) * 1985-10-15 1988-02-16 Bio-Rad Laboratories, Inc. High performance electrophoretic mobilization of isoelectrically focused protein zones
US4810638A (en) * 1986-07-24 1989-03-07 Miles Inc. Enzyme-labeled antibody reagent with polyalkyleneglycol linking group
US4911808A (en) * 1986-09-04 1990-03-27 Bio-Rad Laboratories, Inc. Mobilization of focused protein zones by ion introduction
JP2564540B2 (en) * 1987-04-01 1996-12-18 株式会社日立製作所 Immunoassay method
JPS6432170A (en) * 1987-07-29 1989-02-02 Agency Ind Science Techn Production of novel fluorescent labeled antibody
EP0339302B1 (en) * 1988-04-04 1993-12-22 Teijin Limited Reagent system for immunologically assaying complex of human plasminogen activator inhibitor and human tissue plasminogen activator, and assay kit therefor
US5149661A (en) * 1988-06-08 1992-09-22 Sarasep, Inc. Fluid analysis with particulate reagent suspension
US5006210A (en) * 1989-02-06 1991-04-09 Iowa State University Research Foundation, Inc. Means and method for capillary zone electrophoresis with laser-induced indirect fluorescence detection
US5108568A (en) * 1989-07-07 1992-04-28 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Controlled method of reducing electrophoretic mobility of macromolecules, particles or cells
US5156724A (en) * 1990-01-29 1992-10-20 Millipore Corporation Method for analyzing ionic species using capillary electrophoresis
US5128005A (en) * 1990-01-29 1992-07-07 Millipore Corporation Method for separating ionic species using capillary electrophoresis
FR2671407B1 (en) * 1991-01-08 1994-08-05 Europhor Sa METHOD OF ANALYSIS BY CAPILLARY ELECTROPHORESIS WITH FLUORESCENCE DETECTION, AND DEVICES FOR IMPLEMENTING SAME.
US5120413A (en) * 1991-05-31 1992-06-09 Beckman Instruments, Inc. Analysis of samples utilzing capillary electrophoresis
US5221454A (en) * 1992-01-31 1993-06-22 Biometric Imaging Inc. Differential separation assay
US5137609A (en) * 1992-01-31 1992-08-11 Biometric Imaging Inc. Differential separation assay

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