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WO2003102208A3 - Rapid detection of bt-cry toxins - Google Patents

Rapid detection of bt-cry toxins Download PDF

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Publication number
WO2003102208A3
WO2003102208A3 PCT/IN2003/000199 IN0300199W WO03102208A3 WO 2003102208 A3 WO2003102208 A3 WO 2003102208A3 IN 0300199 W IN0300199 W IN 0300199W WO 03102208 A3 WO03102208 A3 WO 03102208A3
Authority
WO
WIPO (PCT)
Prior art keywords
cry
detection
cry2aa
strips
membrane
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/IN2003/000199
Other languages
French (fr)
Other versions
WO2003102208A2 (en
Inventor
Keshav Raj Kranthi
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Indian Council of Agricultural Research
Original Assignee
Indian Council of Agricultural Research
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Indian Council of Agricultural Research filed Critical Indian Council of Agricultural Research
Priority to AU2003249576A priority Critical patent/AU2003249576A1/en
Priority to MXPA04011769A priority patent/MXPA04011769A/en
Publication of WO2003102208A2 publication Critical patent/WO2003102208A2/en
Publication of WO2003102208A3 publication Critical patent/WO2003102208A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/32—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Bacillus (G)
    • G01N2333/325—Bacillus thuringiensis crystal protein (delta-endotoxin)

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present invention is based on the use of 1) two polyclonal IgGs against two Cry toxins, 2) Cry toxin receptors isolated from lepidopteran larvae and 3) manual striping methods to manufacture immunochromatographic strips that are useful in detecting a variety of analytes. Immunochromatographic strips, which facilitate rapid detection of analytes are expensive if made using monoclonal antibodies and cannot be affordable for Indian farmers and extension workers. The main objective of the current method was to simplify the development of the immunochromatographic detection method for Cry (Bt) toxin detection using affinity purified polyclonal antibodies specific to the analyte and also to provide a robust and easy method suitable for manufacture of 'Cry1Ac/Cry1Ab/Cry2Aa/Cry2Ab toxin' detection strips at affordable cost, under Indian conditions. Anti-Bt (anti-Cry1 Ac or anti-Cry2Aa/Ab or both) antibody or Cry receptor proteins are immobilized on a cellulose nitrate membrane by manual striping. The membrane is blocked using casein or BSA and preservatives. Anti-Bt (anti-Cry1 Ac or anti-Cry2Aa/Ab or both) antibody is labeled with gold and adsorbed on glass-fibre membrane which is placed at the bottom the membrane so as to overlap 2mm. Specificity and accuracy of the assay are greatly enhanced due to the use of antigen-affinity and Protein-A affinity purified polyclonal IgG raised in two different animals (goat and rabbit). The schematic diagram of the Cry toxin detection lateral flow strip assembly is shown in a diagram appended herewith. The strips thus made represent rapid, sensitive devices and methods for detecting the presence of Cry1Ac/Cry1Ab/Cry2Aa/Cry2Ab and crystal toxins either from transgenic plants or in Bt-fermented products. The methods and devices have high sensitivity to detect either single or two Cry toxins simultaneously on a single strip. Use of the present methods provide an assay system which involve a minimal number of steps, and yield reliable results even when used by untrained persons.
PCT/IN2003/000199 2002-05-31 2003-05-29 Rapid detection of bt-cry toxins Ceased WO2003102208A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2003249576A AU2003249576A1 (en) 2002-05-31 2003-05-29 Rapid detection of bt-cry toxins
MXPA04011769A MXPA04011769A (en) 2002-05-31 2003-05-29 Rapid detection of bt-cry toxins.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN600/DEL/02 2002-05-31
IN600DE2002 2002-05-31

Publications (2)

Publication Number Publication Date
WO2003102208A2 WO2003102208A2 (en) 2003-12-11
WO2003102208A3 true WO2003102208A3 (en) 2004-03-25

Family

ID=29596804

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2003/000199 Ceased WO2003102208A2 (en) 2002-05-31 2003-05-29 Rapid detection of bt-cry toxins

Country Status (6)

Country Link
KR (1) KR20050026396A (en)
CN (1) CN100403028C (en)
AU (1) AU2003249576A1 (en)
MX (1) MXPA04011769A (en)
WO (1) WO2003102208A2 (en)
ZA (1) ZA200410268B (en)

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4891317B2 (en) * 2005-06-28 2012-03-07 ズィービーエックス・コーポレーション Membrane array and analytical equipment
CN100348616C (en) * 2005-12-12 2007-11-14 中国农业大学 Bt CrylA antibody, and its preparing method and special antigen and use
US9046518B2 (en) * 2009-04-09 2015-06-02 Hitachi Chemical Company, Ltd. Detector and detection method
CN103197075B (en) * 2012-01-16 2014-10-08 华中农业大学 Method for detecting Bt protein in transgenic rice by quantum dot
EP3680000B1 (en) 2013-03-15 2024-03-13 GlaxoSmithKline Intellectual Property Development Limited Methods for purifying antibodies
CN105693856A (en) * 2016-04-25 2016-06-22 江苏省农业科学院 Monoclonal antibody, cell strain secreting monoclonal antibody, preparation method and application
ES2979153T3 (en) 2016-09-07 2024-09-24 Glaxosmithkline Intellectual Property Dev Methods for purifying antibodies
CN106674334B (en) * 2017-02-08 2020-09-01 金陵科技学院 Cry2 Ad-combined cyclic heptapeptide and encoding gene and application thereof
CN108828230B (en) * 2018-06-21 2021-04-30 北京市农林科学院 Method for rapidly detecting transgenic product by nucleic acid chromatography
CN110346569A (en) * 2019-06-28 2019-10-18 安徽恩禾生物技术有限公司 A kind of thymidine kinase chemoluminescence method detection kit and preparation method thereof
CN114280312B (en) * 2020-09-27 2023-09-15 河北特温特生物科技发展有限公司 Whole blood separation membrane for immunofluorescence chromatography detection and preparation method and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5141850A (en) * 1990-02-07 1992-08-25 Hygeia Sciences, Inc. Porous strip form assay device method
US5712172A (en) * 1995-05-18 1998-01-27 Wyntek Diagnostics, Inc. One step immunochromatographic device and method of use
US6156573A (en) * 1996-11-20 2000-12-05 Monsanto Company Hybrid Bacillus thuringiensis δ-endotoxins with novel broad-spectrum insecticidal activity
WO2001044779A2 (en) * 1999-12-14 2001-06-21 Strategic Diagnostics, Inc. Method of processing and testing powdered samples using immunochromatographic strip tests

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5712171A (en) * 1995-01-20 1998-01-27 Arqule, Inc. Method of generating a plurality of chemical compounds in a spatially arranged array
ES2229395T3 (en) * 1996-11-20 2005-04-16 Monsanto Technology Llc DELTA SPECTRO DELTA ENDOTOXINS.

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5141850A (en) * 1990-02-07 1992-08-25 Hygeia Sciences, Inc. Porous strip form assay device method
US5712172A (en) * 1995-05-18 1998-01-27 Wyntek Diagnostics, Inc. One step immunochromatographic device and method of use
US6156573A (en) * 1996-11-20 2000-12-05 Monsanto Company Hybrid Bacillus thuringiensis δ-endotoxins with novel broad-spectrum insecticidal activity
WO2001044779A2 (en) * 1999-12-14 2001-06-21 Strategic Diagnostics, Inc. Method of processing and testing powdered samples using immunochromatographic strip tests

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AGDIA MOLECULAR DIAGNOSTICS, Catalogue, ImmunoStrip test system for Bt-CrylAb/lAc proteins; ImmunoStrip test system for Bt-Cry3A protein from, http://www.agdia.com (Feb2001) in conjunction with http://web.archive.org. *
ENVIROLOGIX INC., Lateral Flow QuickStix Strip Kit, from http://envirologix.com *
Jenkins, J.L. and Dean D.H. (2001) BMC Biochemistry, vol. 2:12. "Binding specificity of Bacillus thuringiensis CrylAa for purified native Bombyx mori aminopeptidase N and cadherin-like receptors". *
Saxena, D. et al. (1999) Nature, vol. 402, p.480. "Insecticidal toxin in root exudates from Btcorn". *
Stave, J.W. (May/June 2002) Journal of AOAC International, vol. 85(3), pp. 780-786. "Protein immunoassay methods for detection of biotech crops: Applications, limitations and practical considerations". *

Also Published As

Publication number Publication date
AU2003249576A8 (en) 2003-12-19
CN100403028C (en) 2008-07-16
CN1672049A (en) 2005-09-21
ZA200410268B (en) 2006-07-26
KR20050026396A (en) 2005-03-15
AU2003249576A1 (en) 2003-12-19
MXPA04011769A (en) 2005-07-26
WO2003102208A2 (en) 2003-12-11

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