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WO2012046563A1 - Anticorps monoclonal reconnaissant des ciguatoxines ctx1b et un 54-désoxy-ctx1b, et kit de détection de ciguatoxine mettant en œuvre celui-ci - Google Patents

Anticorps monoclonal reconnaissant des ciguatoxines ctx1b et un 54-désoxy-ctx1b, et kit de détection de ciguatoxine mettant en œuvre celui-ci Download PDF

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Publication number
WO2012046563A1
WO2012046563A1 PCT/JP2011/071245 JP2011071245W WO2012046563A1 WO 2012046563 A1 WO2012046563 A1 WO 2012046563A1 JP 2011071245 W JP2011071245 W JP 2011071245W WO 2012046563 A1 WO2012046563 A1 WO 2012046563A1
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Prior art keywords
monoclonal antibody
ctx1b
hybridoma
compound
ciguatoxins
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Japanese (ja)
Inventor
藤井　郁雄
円谷　健
平間　正博
修治 山下
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Osaka Metropolitan University
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Osaka Prefecture University PUC
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/14—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from fungi, algea or lichens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention relates to shigatoxins, particularly a monoclonal antibody capable of recognizing ciguatoxin CTX1B or 54-deoxy-CTX1B having a dihydroxybutenyl group in the A ring, a hybridoma producing the monoclonal antibody, and a shigatoxin using the monoclonal antibody It relates to a kind detection kit.
  • the present invention also relates to a compound used to obtain the above-mentioned monoclonal antibody, and a conjugated compound formed by binding the compound to a carrier protein.
  • Ciguatera toxin is produced by phytoplankton and accumulated in fish and shellfish via the food chain. Therefore, once ciguatera toxin is generated, it becomes a serious social problem because a large number of food fish are toxic in a wide range. Therefore, rapid detection of ciguatera toxin is important for the prevention of food poisoning ciguatera.
  • ciguatoxins which are the major causative toxins of food poisoning ciguatera
  • CTX radioimmunoassay
  • an antibody that recognizes shigatoxins is required.
  • shigatoxins since only a very small amount of shigatoxins is collected from nature (for example, 850 mice, only 0.35 mg of ciguatoxin obtained from 4 tons of moray eel) is difficult to produce by culture, an antibody that recognizes shigatoxins is It is difficult to manufacture.
  • Non-Patent Document 1 Koka et al. Of the University of Hawaii have reported that they produced a conjugate in which ciguatoxin (1 ⁇ g), which is the main body of shigatera toxin, was linked to human serum albumin by the carbodiimide method and used as an antigen to immunize mice to produce a monoclonal antibody.
  • this antibody binds to ciguatoxin, it has also shown a strong cross reaction with okadaic acid, a marine polyether toxin having a similar structure to ciguatoxins. That is, the difference in affinity between this antibody for shigatoxin and okadaic acid is very small (see Non-Patent Document 2).
  • ciguatoxins are not single compounds. That is, ciguatoxins are a mixture of various toxins and have the following formula (III): Four types (compounds 1 to 4) represented by are mainly known.
  • the present inventors previously chemically synthesized the ABC ring portion (partial structure on the left side of Formula (III)) of CTX 1 B (compound 4 in Formula (III) above) of ciguatoxin, and used it as a synthetic hapten.
  • Three monoclonal antibodies were prepared using protein conjugates. However, all of these antibodies showed very weak affinity for ciguatoxin (see Non-patent Document 4).
  • Other research groups have also attempted to immunize animals using a conjugate of a synthetic hapten JKLM ring (partial structure on the right in Formula (III)) to produce a polyclonal antibody that recognizes ciguatoxin. However, no monoclonal antibody has been obtained (see Non-patent Document 5).
  • the present inventors designed and synthesized a synthetic hapten including the IJ KLM ring portion (partial structure on the right side of formula (III)) of ciguatoxin CTX 3 C (compound 1 in formula (III) above)
  • a synthetic hapten including the IJ KLM ring portion (partial structure on the right side of formula (III)) of ciguatoxin CTX 3 C (compound 1 in formula (III) above)
  • hybridoma 3D11 of Accession No. FERM BP-8293 is produced, and monoclonal antibody 3D11 highly specific to shigatoxins is produced using this hybridoma.
  • the dissociation constant (K d ) of this monoclonal antibody 3D11 for shigatoxin CTX3C was 122 nM.
  • the present inventors have designed and synthesized a compound containing the ABCDE ring part (the partial structure on the left side of Formula (III)) of ciguatoxin CTX3C as a hapten, and immunize a mouse with a protein conjugate of this synthetic hapten.
  • the hybridoma 10C9 of Accession No. FERM BP-8292 was produced by the method including, and it succeeded in preparing a monoclonal antibody 10C9 highly specific to shigatoxins using the hybridoma (see Patent Document 2).
  • the present inventors combined the above two types of monoclonal antibodies (3D11 and 10C9) to produce a kit for detecting sigatoxin CTX3C by a sandwich method with further improved detection characteristics (Patent Document 3 and Non-patent Document 3) 6).
  • the present inventors have synthesized, for the first time, a compound having the structure of the HIJKLM ring of ciguatoxins, in order to produce an antibody for detecting ciguatoxin 51-OH-CTX3C and CTX1B having a hydroxy group in the M ring.
  • This is used as a hapten to immunize an animal to prepare hybridoma 8H4 deposited under Accession No. FERM BP-11400, which is used to specifically bind to shigatoxins 51-OH-CTX3C and CTX1B with high specificity.
  • the present inventors have prepared a kit for detecting ciguatoxin 51-OH-CTX3C by a sandwich method having improved detection characteristics by combining the above-mentioned monoclonal antibodies 8H4 and 10C9 (Patent Document 4 and Non-patent Document 7) ).
  • shigatoxin which is the main toxin of ciguatera food poisoning
  • ciguatoxin CTX1B is considered to be the most universally existing ciguatera causative toxin in the world, so its detection is important.
  • the present invention aims to provide means capable of detecting ciguatoxins, particularly ciguatoxins CTX1B and 54-deoxy-CTX1B having a dihydroxybutenyl group in the A ring portion.
  • the present inventors specifically synthesize a compound having the structure of the ABCDE ring part of ciguatoxins having a dihydroxybutenyl group in the A ring part and use it as a hapten to make the compound specific. It has been found that it is possible to obtain a monoclonal antibody that recognizes
  • the present invention provides a monoclonal antibody that specifically recognizes ciguatoxins having a dihydroxybutenyl group in the A ring portion.
  • the present invention is the Accession No. FERM BP-11401 producing the above-mentioned monoclonal antibody on Sep.
  • hybridoma 3G8 (herein, this hybridoma is denoted as "3G8" which is the same name as an antibody) is provided which has been deposited at Tsukuba 1-chome, 1-chome, 1st-address, 1st-center, 6th.
  • the present invention relates to the monoclonal antibody described above and Accession No. FERM BP-11400 on Dec. 22, 2004 National Institute of Advanced Industrial Science and Technology, International Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology (Postal Code 305-8566, Tsukuba, Ibaraki Prefecture, Japan) City, 1-chome, 1-chome, 1st, 1st, 1st, 6th center) 6) The monoclonal antibody produced by hybridoma 8H4, or Accession No.
  • FERM BP-8293 on March 5, 2002, National Institute of Advanced Industrial Science and Technology
  • the present invention provides a sigatoxins detection kit comprising a monoclonal antibody produced by hybridoma 3D11 deposited at the Patent Organism Depositary Center (Postal Code 305-8566, Central 1st, 1-1, 1-chome Higashi, Tsukuba, Ibaraki, Japan).
  • the present invention provides a compound of the following formula (I) in which a compound of the above formula (II) is bound to a carrier protein: (Wherein, n represents an integer) To provide a protein conjugate represented by
  • a system for detecting ciguatoxins which could not be detected by immunological techniques so far particularly ciguatoxins CTX1B and 54-deoxy-CTX1B having a dihydroxybutenyl group in the A ring portion is constructed. be able to. Therefore, according to the present invention, more precise detection of ciguatoxins is possible, and more effective prevention of food poisoning ciguatera is expected.
  • FIG. 1 is a graph showing the antibody titer of mouse serum obtained by immunizing mice with a keyhole limpet hemocyanin (KLH) conjugate, which was evaluated using the BSA conjugate according to the present invention.
  • FIG. 2 is a graph showing the detection characteristics of ciguatoxin CTX1B of the ciguatoxin detection kit according to the present invention.
  • the monoclonal antibody of the present invention is a monoclonal antibody that specifically recognizes ciguatoxins having a dihydroxybutenyl group in the A ring part in the above general formula (III).
  • ciguatoxins having a dihydroxybutenyl group in the A ring portion include ciguatoxin CTX1B (compound 4 in the above formula (III)), shigatoxin 54-deoxy-CTX1B (compound 3 in the above formula (III)) and the like .
  • the monoclonal antibodies of the present invention have K d of about 1 to 100 nM against ciguatoxins CTX1B and 54-deoxy-CTX1B.
  • the monoclonal antibody of the present invention can be obtained by immunizing an animal with the protein conjugate of the above-mentioned formula (I).
  • Known methods can be used to immunize animals.
  • the animals to be immunized include, for example, goats, rabbits and mice, and among them, mice are preferably used.
  • the monoclonal antibody of the present invention is considered to recognize and bind to the ABCDE ring of sigatoxins having a dihydroxybutenyl group in the A ring.
  • the monoclonal antibody of the present invention alone can detect ciguatoxins in an antigen-antibody reaction known per se.
  • the monoclonal antibody of the present invention preferred is, for example, a monoclonal antibody produced by hybridoma 3G8 deposited on Sep. 15, 2010 as the Accession No. FERM BP-11401 at the Patent Microorganisms Depositary, National Institute of Advanced Industrial Science and Technology. It can be used.
  • Shiga toxins are detected with higher sensitivity by using another monoclonal antibody that recognizes another site different from the site of shigatoxins recognized by the monoclonal antibody of the present invention in combination with the monoclonal antibody of the present invention. It can also be done. Therefore, a ciguatoxins detection kit comprising the monoclonal antibody of the present invention and another monoclonal antibody that recognizes a site other than the site of ciguatoxins recognized by the monoclonal antibody is also one of the present invention.
  • monoclonal antibody 8H4 produced by hybridoma 8H4 deposited on December 22, 2004 as a deposit number of FERM BP-11400 at National Institute of Advanced Industrial Science and Technology, National Institute of Advanced Industrial Science and Technology, or Accession Number As the FERM BP-8293, a monoclonal antibody 3D11 produced by hybridoma 3D11 deposited on March 5, 2002 at the National Institute of Advanced Industrial Science and Technology, Patent Microorganisms Depositary on March 5, 2002 can be preferably used.
  • the monoclonal antibody 8H4 is a monoclonal antibody which can be specifically bound to sigatoxins having a hydroxy group in the M ring by using the HIJKLM ring of ciguatoxins having a hydroxy group in the M ring as the synthetic hapten by the present inventors. It was prepared as an antibody (see JP-A-2006-193485).
  • the monoclonal antibody 3D11 is specifically directed to shigatoxins having no hydroxy group in the M ring, by the present inventors using the IJKLM ring of the ciguatoxins having no hydroxy group in the M ring as a synthetic hapten. It was prepared as a monoclonal antibody capable of binding (see Japanese Patent Laid-Open No. 2003-55400).
  • the two types of antibodies that specifically recognize different sites of ciguatoxins as described above can be used in conventionally known immunological methods to detect ciguatoxins.
  • Conventionally known immunological methods include a sandwich method of detecting a target antigen using two or more types of antibodies.
  • the label may be any label commonly used in immunological methods, such as enzyme label (eg, peroxidase, alkaline phosphatase, ⁇ -galactosidase etc.), fluorescent label (eg, fluorescein isothiocyanate (FITC), allophycocyanin (APC) , Phycoerythrin (PE), carbocyanine, etc., radioactive labels (eg, tritium ( 3 H), 125 I, 131 I etc.), gold colloid labels, etc.
  • enzyme labels eg, peroxidase, alkaline phosphatase, ⁇ -galactosidase etc.
  • fluorescent label eg, fluorescein isothiocyanate (FITC), allophycocyanin (APC) , Phycoerythrin (PE), carbocyanine, etc.
  • radioactive labels eg, tritium ( 3 H), 125 I, 131 I etc.
  • gold colloid labels etc.
  • enzyme labels are
  • the compounds of formula (I) above can be obtained from the compounds of formula (II) above.
  • the compound of the formula (I) is prepared by reacting the carrier protein with 3,3'-dithiobis (sulfosuccinimide propionate) (DTSSP) and then tris (2-carboxyethyl) phosphine hydrochloride ( Method of reducing amino group of basic amino acid residue (lysine, histidine and arginine residue) of carrier protein to thiol group by reduction with a reducing agent such as TCEP) and reacting it with a compound of formula (II) It can be obtained by Conventionally, as a method of conjugating a compound containing a ABCDE ring part of ciguatoxins having no dihydroxybutenyl group in the A ring part with a carrier protein, the amino group of the basic amino acid residue of the carrier protein is Methods have been known to react with N-hydroxysuccinimidyl ester derivatives of compounds containing (see Patent
  • any protein may be used as long as it can be used as a carrier to be bound to a hapten in an immunological method, and bovine serum albumin (BSA), keyhole limpet hemocyanin (KLH), egg albumin (OVA), Fibrinogen and the like can be mentioned, and among them, BSA, KLH and OVA are preferable.
  • BSA bovine serum albumin
  • KLH keyhole limpet hemocyanin
  • OVA egg albumin
  • Fibrinogen Fibrinogen and the like
  • Fmoc represents (9-fluorenylmethyloxycarbonyl)
  • DCC represents dicyclohexylcarbodiimide
  • DMAP represents N, N-dimethyl-4-aminopyridine
  • DDQ represents 2,3-dichloro- Represents 5,6-dicyano-p-benzoquinone
  • PPTS represents pyridinium p-toluenesulfonate
  • MeOH represents methanol
  • THF represents tetrahydrofuran
  • DMF represents N, N-dimethylformamide
  • BMPS is 3- 3- N-succinimidyl maleimide is represented
  • NAP represents (naphthylmethyl).
  • DDQ (12.3 mg, 54.0 ⁇ mol) was added to a solution of compound 6 (6.5 mg, 5.4 ⁇ mol) in CH 2 Cl 2 / H 2 O (volume ratio 2: 1, 5.4 mL) at room temperature, and stirred for 30 minutes . After adding a saturated aqueous solution of Na 2 S 2 O 3 to the reaction solution, the reaction solution was diluted with ethyl acetate and then a saturated aqueous solution of NaHCO 3 was added. After extraction with ethyl acetate, the separated organic phase was dried over MgSO 4 .
  • KLH Keyhole Limpet Hemocyanin
  • BSA Bovine Serum Albumin Conjugate: To a solution of BSA (20.0 mg) in PBS (10 mL), DTSSP (13.7 mg, 45.0 ⁇ mol) was added at room temperature. After standing at room temperature for 3 hours, it was dialyzed against PBS (1 L) at 4 ° C. The PBS (1 L) was exchanged twice every two hours, and after 12 hours, the solution was removed from the dialysis membrane to obtain the above compound 12. To a solution of compound 12 in PBS (9.0 mL, 1.70 mg / mL) was added a solution of TCEP (1.8 mg, 6.4 ⁇ mol) in PBS (100 ⁇ L) at room temperature.
  • TCEP 1.8 mg, 6.4 ⁇ mol
  • the BSA conjugate obtained as described above was analyzed by MALDI-TOF-MS.
  • the average molecular weight of the BSA conjugate was about 77400 (the molecular weight of BSA is about 66400). Since the molecular weight of the hapten moiety which is a moiety derived from the compound of the above-mentioned formula (II) is 802, it is understood that an average of 14 haptens are linked to the BSA conjugate.
  • mouse serum was collected, and the antibody titer of the serum was determined by the following ELISA (enzyme-linked immunosorbent assay) method using a conjugate of the compound of formula (II) described above and BSA as an antigen. It was determined.
  • ELISA enzyme-linked immunosorbent assay
  • serum-free medium [RPMI 1640 medium (GIBCO, 11875-085) in 1000 mL penicillin / streptomycin (GIBCO, 15140-122) 10 Medium to which mL was added]
  • the medium was transferred to a 15 mL petri dish, and cells in the spleen were suspended with tweezers.
  • the spleen cell suspension was filtered and transferred to a 50 mL centrifuge tube.
  • 15 mL of serum-free medium was added, pipetting was carried out well, and filtration was performed to a total volume of 30 mL. Centrifuge at 800 rpm for 5 minutes at room temperature, remove the supernatant and tap.
  • a tube in which myeloma cells P3X63-Ag8.653 (Dainippon Pharmaceutical Co., Ltd.) were frozen was taken out from a liquid nitrogen cell storage container at -180 ° C, and thawed quickly in a thermostat at 37 ° C. After thoroughly disinfecting the tube with alcohol cotton, the cell suspension in the tube was transferred to 30 mL of serum-free medium. The supernatant was removed by centrifugation at 800 rpm for 5 minutes at room temperature.
  • growth medium serum free from serum, 100 mL of FBS, 10 mL of L-glutamine (200 mM, GIBCO, 25030-081), 20 mL of Brei clone (BR-001 manufactured by BioResearch Ireland) was added. Medium 10 mL was added, cells were suspended, and transferred to a 50 mL culture flask. The stopper of the flask was loosened, placed in a carbon dioxide incubator, and cultured. The cells were subcultured every 2 to 3 days to make two 250 mL flasks (90 to 100 mL).
  • ECF buffer buffer containing 10 ml of 10 mM magnesium chloride, 10 mL of 10 mM magnesium chloride, 10 mL of 20 mM Tris-HCl (pH 7.2) dissolved in distilled water to make 1000 mL
  • centrifuging 800 rpm, 5 minutes, room temperature
  • the supernatant and tapping twice the operation was repeated with 4.8 mL of ECF buffer.
  • the obtained hybridoma cells (Mouse-Mouse hybridoma) were added to HAT medium [serum free medium 144 mL with FCS 40 mL, L-glutamine (200 mM, 25030-081 manufactured by GIBCO) 2 mL, Brei clone (BioResearch Ireland BR) -001)
  • HAT medium serum free medium 144 mL with FCS 40 mL, L-glutamine (200 mM, 25030-081 manufactured by GIBCO) 2 mL, Brei clone (BioResearch Ireland BR) -001
  • Medium containing 10 mL and HAT (31062-011 manufactured by GIBCO) 4mL was suspended in 60 mL, and 100 ⁇ L / well was transferred to six 96-well plates.
  • a hybridoma which produces an antibody which is cultured for about 2 weeks in a carbon dioxide gas culture apparatus and which binds a conjugate of a compound of formula (II) and BSA (that is, a compound of BSA in the protein in formula (I)) is The conjugate was screened by ELISA. Positive wells were selected, and after cloning twice, hybridomas producing monoclonal antibodies that became ELISA positive were successively cultured and grown to about 200 mL each. Thus, hybridoma 3G8 producing a monoclonal antibody at high antibody titer was obtained.
  • This hybridoma was deposited at National Institute of Advanced Industrial Science and Technology Patent Organism Depositary on Sep. 15, 2010 under Accession No. FERM BP-11401 (hereinafter, this hybridoma is referred to as "3G8" which is the same name as antibody. indicate).
  • the absorbance at 490 nm was measured using a microplate absorbance measurement apparatus (manufactured by BIO-RAD, model 680) to obtain a titration curve.
  • a microplate absorbance measurement apparatus manufactured by BIO-RAD, model 680
  • the above-mentioned HRP-labeled monoclonal antibody 8H4 solution (20 ⁇ g / mL, 50 ⁇ L) was added and allowed to stand at room temperature for 1 hour. Discard the solution and wash 3 times with PBS-Tween, then 100 ⁇ L of substrate solution [Composition of substrate solution: 1,2-phenylenediamine 4.0 mg, hydrogen peroxide solution 10 ⁇ L, 0.1 M citric acid buffer (pH 5.0) 10 mL ] was added, and the color reaction was allowed to proceed for several minutes, and then the reaction was stopped with 2N sulfuric acid (50 ⁇ L). The absorbance at 490 nm was measured using a microplate absorbance measurement device (manufactured by BIO-RAD, model 680). The measurement results are shown in FIG.

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Abstract

La présente invention concerne un anticorps monoclonal permettant de reconnaître une ciguatoxine possédant un groupe dihydroxy-butényle dans un cycle A, un hybridome produisant ledit anticorps monoclonal, et un kit de détection de ciguatoxine. En outre, l'invention concerne un composé mis en œuvre afin d'obtenir ledit anticorps monoclonal, et un composé conjugué constitué par la liaison dudit composé sur une protéine-support.
PCT/JP2011/071245 2010-10-06 2011-09-16 Anticorps monoclonal reconnaissant des ciguatoxines ctx1b et un 54-désoxy-ctx1b, et kit de détection de ciguatoxine mettant en œuvre celui-ci Ceased WO2012046563A1 (fr)

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JP2010226734A JP5757497B2 (ja) 2010-10-06 2010-10-06 シガトキシン類ｃｔｘ１ｂおよび５４−デオキシ−ｃｔｘ１ｂを認識するモノクローナル抗体およびそれを用いるシガトキシン類検出キット

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TR201911272T4 (tr) * 2012-08-21 2019-08-21 Janssen Pharmaceutica Nv Paliperidon haptenleri.
WO2014031587A1 (fr) * 2012-08-21 2014-02-27 Janssen Pharmaceutica Nv Haptènes de l'olanzipine
AU2013305935B2 (en) * 2012-08-21 2017-08-24 Saladax Biomedical Inc. Haptens of quetiapine for use in immunoassays

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000001417A1 (fr) * 1998-07-03 2000-01-13 Cyclacel Limited Systeme d'administration
WO2000029427A2 (fr) * 1998-11-13 2000-05-25 Cyclacel Limited Vecteurs de transport
JP2003267978A (ja) * 2002-03-12 2003-09-25 Japan Science & Technology Corp Ｃｔｘ３ｃのａ−ｅ環部を持つハプテン部を有するタンパク質コンジュゲートで免疫して得られるシガトキシンｃｔｘ３ｃ抗体およびその作製方法
JP2003267979A (ja) * 2002-03-12 2003-09-25 Japan Science & Technology Corp シガトキシンｃｔｘ３ｃを検出するサンドイッチ測定キット類
JP2006193485A (ja) * 2005-01-14 2006-07-27 Osaka Prefecture シガトキシン類を認識するモノクローナル抗体、およびそれを用いるシガトキシン類検出キット
WO2007083638A1 (fr) * 2006-01-19 2007-07-26 Japan Science And Technology Agency Méthode de synthèse de ciguatoxine ctx1b et composé utile dans la synthèse de ciguatoxine ctx1b
JP2011200164A (ja) * 2010-03-25 2011-10-13 Osaka Prefecture Univ シガトキシン類を認識するヒト化抗体

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000001417A1 (fr) * 1998-07-03 2000-01-13 Cyclacel Limited Systeme d'administration
WO2000029427A2 (fr) * 1998-11-13 2000-05-25 Cyclacel Limited Vecteurs de transport
JP2003267978A (ja) * 2002-03-12 2003-09-25 Japan Science & Technology Corp Ｃｔｘ３ｃのａ−ｅ環部を持つハプテン部を有するタンパク質コンジュゲートで免疫して得られるシガトキシンｃｔｘ３ｃ抗体およびその作製方法
JP2003267979A (ja) * 2002-03-12 2003-09-25 Japan Science & Technology Corp シガトキシンｃｔｘ３ｃを検出するサンドイッチ測定キット類
JP2006193485A (ja) * 2005-01-14 2006-07-27 Osaka Prefecture シガトキシン類を認識するモノクローナル抗体、およびそれを用いるシガトキシン類検出キット
WO2007083638A1 (fr) * 2006-01-19 2007-07-26 Japan Science And Technology Agency Méthode de synthèse de ciguatoxine ctx1b et composé utile dans la synthèse de ciguatoxine ctx1b
JP2011200164A (ja) * 2010-03-25 2011-10-13 Osaka Prefecture Univ シガトキシン類を認識するヒト化抗体

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
GHOSH SS. ET AL.: "Use of maleimide-thiol coupling chemistry for efficient syntheses of oligonucleotide-enzyme conjugate hybridization probes", BIOCONJUGATE CHEM., vol. 1, 1990, pages 71 - 76, XP002131636, DOI: doi:10.1021/bc00001a009 *
KATSUTOSHI TAKEUCHI ET AL.: "Ciguatoxin CTX1B ni Ketsugo suru Kotai Sakusei o Mokuteki to shita Hapten - Tanpakushitsu Conjugate no Gosei", 91ST ANNUAL MEETING OF CHEMICAL SOCIETY OF JAPAN IN SPRING (2011) KOEN YOKOSHU IV, 11 March 2011 (2011-03-11), pages 1134 *
OGURI H.: "Bioorganic Studies Utilizing Rationally Designed Synthetic Molecules: Absolute Configuration of Ciguatoxin and Development of Immunoassay Systems", BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN, vol. 80, no. 10, 2007, pages 1870 - 1883 *
SHUJI YAMASHITA ET AL.: "A New Convergent Synthesis of Left Wing Fragment of Ciguatoxin CTX1B", 86TH ANNUAL MEETING OF CHEMICAL SOCIETY OF JAPAN IN SPRING (2006) KOEN YOKOSHU II, 13 March 2006 (2006-03-13), pages 1365 *
SHUJI YAMASHITA ET AL.: "Ciguatoxin CTX1B Hidarigawa Fragment no Gosei Kenkyu", 87TH ANNUAL MEETING OF CHEMICAL SOCIETY OF JAPAN IN SPRING (2007) KOEN YOKOSHU II, 12 March 2007 (2007-03-12), pages 1208 *

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10890573B2 (en) 2017-12-19 2021-01-12 International Business Machines Corporation Facile methods to detect toxin in seafood

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