GB2466536A - Immunoaffinity column for detection of deoxynivalenol, deoxynivalenol, T2 toxin and HT2 toxin - Google Patents

Immunoaffinity column for detection of deoxynivalenol, deoxynivalenol, T2 toxin and HT2 toxin Download PDF

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Publication number
GB2466536A
GB2466536A GB0914801A GB0914801A GB2466536A GB 2466536 A GB2466536 A GB 2466536A GB 0914801 A GB0914801 A GB 0914801A GB 0914801 A GB0914801 A GB 0914801A GB 2466536 A GB2466536 A GB 2466536A
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Prior art keywords
column
toxin
deoxynivalenol
resin
zearalenone
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GB0914801D0 (en
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Simon James Bevis
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    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56961—Plant cells or fungi
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
    • B01D15/3804—Affinity chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
    • B01D15/3804—Affinity chromatography
    • B01D15/3809—Affinity chromatography of the antigen-antibody type, e.g. protein A, G, L chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28052—Several layers of identical or different sorbents stacked in a housing, e.g. in a column
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
    • B01J20/3206—Organic carriers, supports or substrates
    • B01J20/3208—Polymeric carriers, supports or substrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3268—Macromolecular compounds
    • B01J20/3272—Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
    • B01J20/3274—Proteins, nucleic acids, polysaccharides, antibodies or antigens
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/545—Synthetic resin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
    • B01J2220/54—Sorbents specially adapted for analytical or investigative chromatography
    • 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
    • G01N2333/37—Assays involving biological materials from specific organisms or of a specific nature from fungi

Abstract

A multi analyte immunoaffinity column is disclosed comprising at least one unit of resin having deoxynivalenol specific affinity and, for each unit of resin having deoxynivalenol specific affinity, the column further contains about 0.4 - 0.6 units of resin containing antibody having specificity for Zearalenone and about 0.4 - 0.6 units of resin containing antibody having specificity for T2 toxin and HT2 toxin. One unit of resin is the quantity of resin containing antibody that will bind 750ng of Deoxynivalenol, 875ng of Zearalenone and 1300ng of T2 and HT2 toxin, respectively. A method of analyzing a single liquid sample for these toxins using the aforementioned multi analyte column is also claimed.

Description

UK patent application Immunoaffinity column for detection of Deoxynivalenol, Zearalenone, 12 toxin and HT2 toxin A multi analyte immunoaffinity column is disclosed. The column may contain at least one unit of resin having deoxynivalenol specific affinity and, for each unit of resin having deoxynivalenol specific affinity, the column further contains about 0.4-0.6 units of resin containing antibody having specificity for Zearalenone and about 0.4 0.6 units of resin containing antibody having specificity for 12 toxin and HT2 toxin. One unit of resin is the quantity of resin containing antibody that will bind 7SOng of Deoxynivalenol, 875ng of Zearalenone and l300ng of 12 and HT2 toxin.
Description
Field of the invention
The invention is concerned with an affinity column used for immunological screening for environmentally occurring toxins, for example, those found in food products, and is particularly directed to multi analyte columns for detecting a plurality of toxins that may be present in a single sample.
Background of the invention
Awareness of the incidence and effect of human and animal exposure to toxins by humans and other animals via food, water and air is of critical importance to our survival. The detection of toxins such as Deoxynivalenol, Zearalenone, T2 and HT2 has become especially important. In particular screening procedures for assessing the exposure of humans to such toxins may require the ability to quantify both the toxin and its metabolites.
lmmunoaffinity columns for the detection of single analytes, for example, one of deoxynivalenol, Zearalenone, 12 or HT2 in a sample are well known; (Yang et al, Aspects of Applied Biology 68, 2003, Schumacher et al Fresnius J Analytical Chem (1998) 360: 241-245, Heidtmann et al poster publication 2007), however columns targeting higher numbers of chemical species necessarily must capture more diverse analytes. Deoxynivalenol is a highly polar toxin that is smaller than a molecule of table sugar sucrose with a molecular weight of 296.3, Zearalenone is an estrogenic resorcyclic acid lactone compound with a molecular weight of 318.4 and T2 and HT2 toxins are closely related epoxy sesquiterpenoids with respective molecular weights of 466.5 and 424.5. Thus the preparation of multi analyte columns and their methods of use increase in complexity out of proportion to the number of toxins being added for analysis.
Column development must allow for treatment of all target analytes according to similar methods in order that they all be analysed with a single column.
There have been several examples of simultaneous natural occurrence of Deoxynivalenol, Zearalenone, T2 and HT2 in various substrates. Human food and animal feed (particularly in cereal based products) can contain different multi toxin combinations from fungi infected grains. A desire still exists for competent multi analyte columns for analysing a plurality of toxins with a single column.
Summary of the invention
The present invention provides a multi analyte immunoaffinity column capable of analysing a single sample containing one or more of Deoxynivalenol, Zearalenone, T2 and RT2. The multianalyte columns in accord with the present invention comprise a first quantity of a first resin comprising an antibody having specificity for Deoxynivalenol, a second quantity of a second resin comprising an antibody having specificity for Zearalenone, and a third quantity of a third resin comprising an antibody having specificity for T2 and HT2.
It is desirable to obtain at least a 60%, preferably at least a 70% recovery from the column for each toxin in the sample. It is also desirable to have a column flow rate of approximately 2-3ml per minute, preferably so that a 25m1 sample will flow through the column in approximately 10 minutes. We have found that it is not possible to obtain satisfactory analytical results in a multi analyte column by merely combining the quantities of resin used in a single analyte column to analyse each particular analyte.
Thus we have found that a multi analyte column capable of analysing a single sample containing Deoxynivalenol, Zearalenone, T2 toxin and HT2 toxin comprises for each unit of resin containing antibody having specificity for Deoxynivalenol about 0.4 -0.6 units of resin containing antibody having specificity for Zearalenone and about 0.4-0.6 units of resin containing antibody having specificity for T2 and HT2 toxins, As used herein one unit of resin is defined as the quantity of resin containing antibody that will bind 7SOng of Deoxynivalenol, 875ng of Zearalenone, 1,300ng of T2 toxin and HT2 toxin, respectively.
In a preferred embodiment, the multi analyte column of the present invention is capable 0f analysing a sample to detect Deoxynivalenol, Zearalenone, T2 and HT2 in the analysis of a single sample applied to the column.
The invention also provides a method for analysing a single sample for Deoxynivalenol, Zearalenone, T2 toxin and HT2 toxin, the method comprising providing a multi analyte column as described herein, applying liquid sample suspected of containing one or more of the specified toxins to bind any of the specified toxins to resins in the column, washing the column, eluting the resins and analysing the eluant for the presence of each of the specified toxins. The liquid sample can be a liquid suspected of containing toxins or a liquid extract of a solid material suspected of containing toxins. Specific examples of sample materials that can be analysed in accord with the columns of the present invention include fungi infected grains for example wheat, barley, oats, beverages, for example beer, malt based drinks, animal feed and baby food.

Claims (11)

  1. Claims 1) A multi analyte column comprising at least 1 unit of resin having deoxynivalenol specific affinity and, for each unit of resin having deoxynivalenol specific affinity, the column further contains about 0.4-0.6 units of resin containing antibody having specificity for Zearalenone and about 0.4- 0.6 units of resin containing antibody having specificity for T2 toxin and HT2 toxin. Wherein one unit of resin is the quantity of resin containing antibody that will bind 750ng of Deoxynivalenol, 875ng of Zearalenone and l300ng of T2 and HT2 toxin respectively.
  2. 2) The multianalyte column of claim 1, wherein the column is structured and arranged to recover at least 60% and preferaby at least 70% of a toxin in a 25m1 sample being analysed, said toxin being selected from the group consisting of Deoxynivalenol, Zearalenone, T2 and HT2.
  3. 3) The multianalyte column of claim 1, wherein the column is structured and arranged to have a column flow rate of approximately 2-3m1 /minute, and or wherein a flow rate of sample fluid through the column is 1 -2 drops per second.
  4. 4) The multi analyte column of claim 1, wherein the column is capable of analysing a sample to detect each of Deoxynivalenol, Zearalenone, 12 and HT2.
  5. 5) The multi analyte column of claim 1, wherein the resin has a particle size of about 45-165 micron.
  6. 6) The niultianalyte column of any one of the preceding claims wherein one resin, having a toxin specific affinity is layered into a column followed successively by layering into the column another resin having a different toxin specific affinity until all of the resin is in the column.
  7. 7) The multianalyte of any one of claims 1-5, wherein the resins having different toxin specific affinity are mixed and placed in a column.
  8. 8) A method of analysing a single liquid sample for Deoxynivalenol, Zearalenone, T2 and HT2, the method comprising: providing a multi analyte column comprising at least one unit of resin having deoxynivalenol specific affinity and the column further contains about 0.4-0.6 units of resin containing antibody having specificity for Zearalenone, about 0.4 -0.6 units of resin containing antibody having specificity for T2 toxin and HT2 toxin. Wherein, one unit of resin is the quantity of resin containing antibody that will bind 750ng of Deoxynivalenol, 875ng of Zearalenone and l300ng of 12 and HT2 toxin respectively; loading the column with a predetermined amount of a liquid or fluid sample suspected of containing one or more of the toxins selected from Deoxynivalenol, Zearalenone, 12 and HTZ; binding the toxins to the antibodies on the column; subsequently eluting each of the toxins in eluant; and analysing the eluant for the presence of each toxin.
  9. 9) The method according to claim 8, wherein toxins are extracted from a food using a water based or water compatible solvent.
  10. 10) The method according to claim 9, wherein the solvent is water:methanol, water:acetonitrile, ethanol, water:ethanol, a salt solution or a buffer solution.
  11. 11) The method according to any one of claims 8 -10, wherein the liquid or fluid sample comprises a liquid or fluid sample selected from the group consisting of a food extract, a grain extract, an alcoholic beverage, a food product or a component of a food product, a grain to be analysed for a fungi infection, a malt beverage or animal feed or baby food.
GB0914801A 2009-08-25 2009-08-25 Immunoaffinity column for detection of deoxynivalenol, deoxynivalenol, T2 toxin and HT2 toxin Withdrawn GB2466536A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103575885A (en) * 2012-07-19 2014-02-12 北京勤邦生物技术有限公司 Enzyme linked immunoassay kit for detecting T-2 toxin, and application thereof
CN103801110A (en) * 2012-11-07 2014-05-21 江苏维赛科技生物发展有限公司 Preparation method of deoxynivalenol immunoaffinity column
CN103801111A (en) * 2012-11-08 2014-05-21 江苏维赛科技生物发展有限公司 Preparation method for T-2 toxin immunoaffinity column
CN104569382A (en) * 2013-10-26 2015-04-29 北京勤邦生物技术有限公司 Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column
CN110018306A (en) * 2019-03-20 2019-07-16 江苏大学 It is a kind of for quickly detecting the preparation method of the sol-gel immune affinity chromatographic column of deoxynivalenol

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1757349A1 (en) * 2005-08-26 2007-02-28 Vicam, L.P. Multi-analyte affinity column
EP1787699A1 (en) * 2005-11-17 2007-05-23 Vicam, L.P. Multi-analyte affinity column

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1757349A1 (en) * 2005-08-26 2007-02-28 Vicam, L.P. Multi-analyte affinity column
EP1787699A1 (en) * 2005-11-17 2007-05-23 Vicam, L.P. Multi-analyte affinity column

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Food Contaminants: Mycotoxins and Food Allergens, Book Chapter, ACS symposium series, Rudrabhatla Mahalakshmi, et al., 2008, "Detection of Relevant Mycotoxins in Wheat Beer and Sake by LC-MS/MS Using Prototype Immunoaffinity Column Clean-Up: A Preliminary Study" pages 241-251. *
Mycotoxin Factbook: Food & Feed Topics, Book Chapter, M Pascale, et al., 2006, "Immunoaffinity clean-up/fluorescence detection methods for mycotoxins", pages 269-283. *
Rapid communications in mass spectrometry, Vol. 21, No. 20, 2007, V Lattanzio et al., "Simultaneous determination of aflatoxins, ochratoxin A and Fusarium toxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup.", pages 3253-3261. *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103575885A (en) * 2012-07-19 2014-02-12 北京勤邦生物技术有限公司 Enzyme linked immunoassay kit for detecting T-2 toxin, and application thereof
CN103801110A (en) * 2012-11-07 2014-05-21 江苏维赛科技生物发展有限公司 Preparation method of deoxynivalenol immunoaffinity column
CN103801111A (en) * 2012-11-08 2014-05-21 江苏维赛科技生物发展有限公司 Preparation method for T-2 toxin immunoaffinity column
CN104569382A (en) * 2013-10-26 2015-04-29 北京勤邦生物技术有限公司 Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column
CN104569382B (en) * 2013-10-26 2016-09-21 北京勤邦生物技术有限公司 A kind of immune affinity column detecting vomitoxin and application thereof
CN110018306A (en) * 2019-03-20 2019-07-16 江苏大学 It is a kind of for quickly detecting the preparation method of the sol-gel immune affinity chromatographic column of deoxynivalenol

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