WO1990006954A1 - Anticorps monoclonaux reconnaissant les polysaccharides polysulfates - Google Patents

Anticorps monoclonaux reconnaissant les polysaccharides polysulfates Download PDF

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Publication number
WO1990006954A1
WO1990006954A1 PCT/AU1989/000547 AU8900547W WO9006954A1 WO 1990006954 A1 WO1990006954 A1 WO 1990006954A1 AU 8900547 W AU8900547 W AU 8900547W WO 9006954 A1 WO9006954 A1 WO 9006954A1
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monoclonal antibody
polysulphated
antibody
solid support
labelled
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PCT/AU1989/000547
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English (en)
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Peter Ghosh
Prachya Kongtawelert
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The University Of Sydney
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids

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  • the present invention relates to the preparation of monoclonal antibodies (MAb) against polysulphated polysaccharides which contain 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution, to their use in the detection and/or quantification of polysulphated polysaccharides which contain 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution in biological samples and to diagnostic kits for use in the detection and/or quantification of those polysulphated polysaccharides comprising the monoclonal antibodies of the invention.
  • MAb monoclonal antibodies
  • Dextran sulphates have also been used for many years as antilipemic agents (Ninomiya fit .al, 1988) but more recently attention has focused on the ability of a low molecular weight fraction (M 8000) to block the binding of certain virions to T lymphocytes (Baba fit al 1988a; Chang fit S , 1988; Mitsuya fit .il, 1988). This latter activity, has also been exhibited by PPS (Baba fit ⁇ , 1988c) and is currently being explored clinically as a therapeutic means of suppressing the effects of HIV-1 and HIV-2 viruses which are responsible for acquired immune deficiency syndrome (AIDS) in man.
  • AIDS acquired immune deficiency syndrome
  • Enzyme-linked immunosorbant assays are now widely used for the assay of a variety of hormones, drugs, polysaccharides, proteins, and cell derived products, and this technique offers many advantages over other methods.
  • ELISA Enzyme-linked immunosorbant assays
  • polsulphated polysaccharides can have anti-serine protease such as anti-elastase activity. It is recognised that serine proteases such as elastase are involved in inflammatory conditions such as arthritis, destruction of connective tissue and emphysema as well as other inflammatory and degenerative diseases and viral infections. Thus compounds with anti-serine protease activity such as antielastase activity are implicated as therapeutic agents for these conditions.
  • the present inventors have surprisingly found that there is a good correlation between antielastase activity and binding to the monoclonal antibody of the invention for polysulphated polysaccharides.
  • the antibodies of the invention can be of use in the selection of drugs and suggest that monoclonal antibodies may be of use in selection of drugs where a particular conformation and charge density, recognised by a monoclonal is central to the activity of the drug.
  • the present invention relates to the preparation, and characterisation of specific MAbs against polysulphated polysaccharides.
  • Enzyme-linked immunosorbent inhibition assays (ELISIA) were set-up using these antibodies to allow detection and/or quantification of polysulphated polysaccharides in human serum or plasma and/or other biological specimens.
  • ELISIA enzyme-linked immunosorbent inhibition assays
  • pentosan polysulphate and dextran sulphate as examples of the methods developed, we defined the limits for detection of these molecules, and/or then applied the assay to determine the levels of pentosan polysulphate in plasma samples of three healthy volunteers, 30 minutes, 1 hour and 2 hours after IV administration.
  • the method described here offers selectivity, sensitivity and convenience for the quantification of polysulphated polysaccharides in biological fluids.
  • the small study of plasma levels of pentosan polysulphate achieved after i»v. administration of the drug served to illustrate this point, for the results obtained using our assay compared favourably with reports by others using radioactivity-labelled pentosan polysulphate (MacGregor fit al 1984) or a competive binding assay (MacGregor f it al 1985) .
  • our MAb only recognised pentosan polysulphate, its desulpated metabolite xylan, which appears in the circulation within one to two hours of drug administration (MacGregor fit al 1984), was not falsely included during the quantification of the drug levels.
  • the techniques described here thus offer a means of routinely detecting and/or quantifying polysulphated polysaccharides which contain 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution in biological samples and the MAbs when coupled with fluorescent, radioactive materials or chromogens could be used to visualise or localise the presence of polysulphated polysaccharides which contain 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution in histological sections or cell preparations.
  • a monoclonal antibody against a polysulphated polysaccharide containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution Typically the monoclonal, antibody recognises pentosan polysulphate, glycosaminoglycan polysulphates, dextran sulphate, carrageenan and chondroitin sulphate E, but does not significantly react with heparin, heparan sulphate, chondroitin sulphate A, B, C or D or keratan sulphate.
  • a preferred monoclonal antibody of the invention is MAb 5-B-10.
  • a monoclonal antibody of the first embodiment in labelled form.
  • the label is selected from an enzyme, a radioisotope a, fluorescent label or biotin.
  • a method for the detection and/or quantification of a polysulphated polysaccharide in a sample comprises: providing a solid support coated with a polycationic coating material and with a polysulphated polysaccharide containing 2,3-, 2,6 or 4,6- disulphate ester pyranose ring substitution; providing a sample; adding a labelled monoclonal antibody of the second embodiment, to the sample; applying the labelled monoclonal antibody containing sample to the solid support; removing unbound sample; and detecting the label.
  • the solid support is selected from PVC, polystyrene, Sepharose and agarose.
  • the solid support is PVC.
  • the solid support may be formed as a microtitration plate, tube or bead.
  • the polycationic coating material is poly-L-lysine.
  • polysulphated polysaccharide coat is selected from pentosan polysulphate, dextran sulphate, glycosaminoglycan polysulphates, carrageenan and chondroitin sulphate E.
  • polysulphated polysaccharide is pentosan polysulphate or dextran sulphate.
  • monoclonal antibody is MAb 5-B-10.
  • the sample is a serum or plasma sample.
  • a method for the detection and/or quantification of polysulphated polysaccharides containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution in a sample comprises: providing a solid support coated with a polycationic coating material and with a polysulphated polysaccharide containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution; providing a sample; adding a monoclonal antibody of the first embodiment to the sample; applying the monoclonal antibody containing sample to the solid support; removing unbound sample; applying a labelled antibody reactive with the monoclonal antibody to the solid support; removing unbound labelled antibody; and detecting the label.
  • the solid support is selected from PVC, polystyrene, agarose and sepharose.
  • the solid support is PVC.
  • the polycationic coating material is poly -L-lysine.
  • the polysulphated polysaccharide coat is selected from pentosan polysulphate, dextran sulphate, chondroitin sulphate E, carrageenan and glycosaminoglycan polysulphates .
  • polysulphated polysaccharide coat is pentosan polysulphate or dextran sulphate.
  • the monoclonal antibody is MAb-5-B-10.
  • the labelled antibody is an anti-mouse antibody, preferably an anti-mouse IgM antibody.
  • the label is selected from an enzyme, a fluorescent label, a radiolabel or biotin.
  • the labelled antibody is alkaline phosphatase conjugated rabbit anti-mouse IgM.
  • a diagnostic kit for use in the detection and/or quantification of polysulphated polysaccharides containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution which kit comprises: a solid support; a polycationic coating material; a polysulphated polysacharide coating material containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution; and a labelled monoclonal antibody of the second embodiment.
  • the solid support is selected from PVC, polystyrene, agarose and sepharose.
  • the solid support is PVC.
  • the polysulphated polysaccharide coating is selected from pentosan polysulphate, dextran sulphate, chondroitin sulphate E, carrageenan and glycosaminoglycan polysulphates.
  • the polysulphated polysaccharide coating is pentosan polysulphate or dextran sulphate.
  • the labelled monoclonal antibody is MAb 5-B-10.
  • the label is alkaline phosphatase.
  • a diagnostic kit for use in the detection and/or quantification of polysulphated polysaccharides containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution which kit comprises: a solid support; a polycationic coating material; • a polysulphated polysaccharide coating material containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution; a monoclonal antibody of the first embodiment; and a labelled antibody which recognises the monoclonal antibody.
  • the solid support is selected from PVC, polystyrene, agarose and sepharose.
  • the solid support is PVC.
  • the polysulphated polysaccharide coating is selected from pentosan polysulphate, dextran sulphate, glycosaminoglycan polysulphates, carrageenan and chondroitin sulphate E.
  • the polysulphated polysaccharide coating is pentosan polysulphate or dextran sulphate.
  • the monoclonal antibody is MAb 5-B-10.
  • the labelled antibody is selected from anti-mouse immunoglobulin antibodies.
  • the labelled antibody is an alkaline phosphatase labelled rabbit anti-mouse IgM antibody.
  • the solid support is coated with poly-L-lysine.
  • the poly-L-lysine coated support is provided coated with the polysulphated polysaccharide.
  • a method for determining whether a polysulphated polysaccharide is a potential antiarthritic, anti-inflammatory, anti-degenerative, or anti-viral drug comprises: determining whether the polysulphated polysaccharide is recognized by a monoclonal antibody of the first embodiment.
  • the monoclonal antibody is MAb 5-B-10.
  • an eighth embodiment of this invention there is provided a method for the visualisation and/or localisation of polysulphated polysaccharides containing 2,3-, 2,6- or 4,6- disulphate ester pyranose ring substitution in tissues and/or histological specimens, comprising administering a labelled antibody of the second embodiment to the tissue or specimen.
  • Figure 1 shows inhibition curves for the characterization of MAb 5-B-10 using the enzyme-linked immunosorbent- inhibition assay (ELISIA) .
  • Figure 2 shows sub-typing of the MAb 5-B-10 by the ELISA technique employing sub-type specific rabbit anti-mouse immunoglobulins.
  • ELISIA enzyme-linked immunosorbent- inhibition assay
  • Figure 3 shows (a) elution profile of MAb 5-B-10 on
  • Figure 4 shows the saturation curve of MAb 5-B-10 in human serum obtained using the ELISA technique and wells coated with 25 ⁇ g/ml of pentosan polysulphate.
  • Figure 5 shows the titration curve of various concentrations of pentosan polysulphate in PBS, pH 7.4 for coating wells using the ELISA technique with MAb 5-B-10 dilution of 1:400 in human serum.
  • Figure 6 shows a typical standard * inhibition curve obtained for the quantification of pentosan polysulphate
  • Figure 8 shows the kinetics of clearance of pentosan polysulphate (lmg/kg) in plasma of human volunteers given the drug intravenously as measured by the ELISIA technique using MAb 5-B-10.
  • the present invention provides a method for detection and/or quantification of polysulphated polysaccharides in human serum and plasma by developing monoclonal antibodies that recognise the polysulphated polysaccharides and using the monoclonals in an ELISIA in the following way:
  • the absorption at a characteristic wavelength is determined using a microtitration plate reader.
  • the preferred concentration of human sera or other biological fluids containing the polysulphated polysaccharide is 20 - 10,000 ng/ml. If is preferable that this serum be incubated with an equal volume of its monoclonal antibody in polypropylene tubes at room temperature for approximately one hour.
  • the preferred but not exclusive primary substrate is NADP in a diethanolamine buffer.
  • the preferred but not exclusive secondary substrate is alcohol dehydrogenase diaphorase at 2.1 - 2.4 u/ml; p-iodonitrotetrazolium violet absolute ethanol all dissolved in phosphate buffer pH 7.2. It is preferable to stop the reaction by the addition of HC1.
  • Iscove's modified Dulbecco's medium was obtained from Gibco Laboratories, Grand Island, N.Y., U.S.A. Freund's adjuvants were purchased from Difco Laboratories, Detroit, Michigan, U.S.A. Alkaline phosphatase conjugated rabbit anti-mouse immunoglobulins were from Dakopatts, Glostrup, Denmark. Alkaline phosphatase conjugated goat anti-rabbit IgG, rabbit anti-mouse (subclass specific) immunoglobulin panel and a high performance hydroxypatite column were purchased from Bio-Rad Laboratories, Richmond, CA, U.S.A. Xylan, dextran sulphate (M.W.
  • Sodium pentosan polysulphate is the generic chemical name for the polysulphate ester of xylan which is a polymer prepared semisynthetically from plant raw materials.
  • the repeating units of xylan polymer are (1—4) linked ⁇ -D-xylopyranoses, with one molecule of the sulphated esters of ⁇ -D-glucopyranosyluronic acid attached to the 2 position of the xylan every nine monomeric sugars.
  • the degree and positions of substitution of the sulphate esters and ring conformation of pentosan polysulphate have been confirmed by 13C-NMR spectroscopy as well as classical analytical techniques (e.g.
  • mice Female Balb/c mice aged 6-8 weeks were immunized with polysulphated polysaccharide, e.g. pentosan polysulphate (lmg/ml in PBS) mixed with an equal volume of complete Freund's adjuvant. This mixture (50 ⁇ l) was injected into the footpad of both hind legs.
  • polysulphated polysaccharide e.g. pentosan polysulphate (lmg/ml in PBS) mixed with an equal volume of complete Freund's adjuvant. This mixture (50 ⁇ l) was injected into the footpad of both hind legs.
  • cells from the spleen and lymph node of immunized mice were isolated and were fused with the Balb/c mouse myeloma line X63-Ag8.653 (a gift from Dr. M. Pollard, Sutton Research Laboratories, The Royal North Shore Hospital of Sydney) according to the basic principle described by Kohler and Milstein (1975) (Galfre and Milsten 1981).
  • Hybridomas obtained from positive wells were cloned and subcloned by the limiting dilution technique using Balb/c mouse macrophage cells as feeders, grown in bulk culture. Ascitic fluids were then produced by injecting the hybridorna cell line intra-peritoneally to pristane primed Balb/c mice. Monoclonal antibody 5-B-10 specific for the polysulphated polysaccharides, dextran sulphate and pentosan polysulphate was isolated and characterised from the ascitic fluid, as described below. Other monoclonals specific for Arteparon , heparin and heparan sulphate were isolated by the same method.
  • Hybridomas were obtained from the fusion of spleen cells and myelomas using polyethylene glycol as fusing agent. Hybridomas from the highest titer of ELISA-positive wells were expanded and cloned by the limiting dilution technique. We obtained eight clones which displayed significant binding to a pentosan polysulphate coated plate as determined by the ELISA. Clone number 5-B-10 was selected from these to produced ascitic fluids in Balb/c mice. When mice were innoculated with Arteparon , heparin or heparan sulphate other clones were obtained which showed specificity for these sulphated polysaccharides. Here, we describe as an example the results obtained for 5-B-10.
  • the diluted ascitic fluid was successfully characterized by sub-typing of the immunoglobulins by specific antibodies and the ELISA technique.
  • Hybridoma clone 5-B-10 was found to produce IgM immunoglobulins and kappa light chains as shown in Figure 1.
  • Enzvme-linked Immunosorbent Assay for the Detection of MAbs in Tissue Culture Media
  • Alkaline phosphatase substrate (lmg/ml of p-nitrophenyl phosphate in 1.0M diethanolamine and 0.001M MgCl ⁇ ) was then added and the absorption at 405nm which developed at 37°C after 60 minutes incubation was determined using a microtitration plate reader after the reaction was stopped by the addition of 50 ⁇ l of 5N NaOH.
  • ELISIA Immunosorbant-inhibition Assay
  • Ascitic fluids which were obtained from the Balb/c mice were diluted, centrifuged (10,000g;10 minutes) and supernatants applied to a high performance hydroxyapatite column (HPHT with guard column) .
  • Proteins and MAb were eluted from the column by using a linear gradient of 10-350mM phosphate buffer, pH 6.8 at a flow rate of lml/minute collecting 1.0ml fractions. Fractions were monitored for absorption at OD280 and tested for antibody activity by using the ELISA technique as described. Fractions which contained protein and antibody activity were pooled and concentrated by ultrafiltration.
  • the MAb clone 5-B-10 was identified as an IgM immunoglobulin with kappa light chains which would be the expected response of the animal since the fusion was achieved just 17 days after the primary immunization by foot-pad injection of antigen.
  • This particular MAb while interacting with both pentosan polysulphate and dextran sulphate did not cross react with heparin, heparin sulphate, Arteparon , chondroitin sulphates, dermatan sulphate, keratan sulphate, hyaluronic acid or xylan.
  • MAbs specific for heparin, heparin sulphate and Arteparon were obtained. Characterization of MAb 5-B-10 bv ELISIA Technique
  • Figure 2 shows the inhibition curves obtained using MAb 5-B-10 in the ELISIA for the polysaccharides pentosan polysulphate, dextran sulphate, xylan, sodium heparin and Arteparon . No interaction was found for dermatan sulphate, chondroitin -4- or -6- sulphate, keratin sulphate or hyaluronic acid (data not shown) . It is clear that using MAb 5-B-10, only pentosan polysulphate and dextran sulphate were inhibitory to the binding of the antibody to pentosan polysulphate coated wells.
  • Activated polyvinyl chloride immuno-assay plates were pre-coated with poly-L-lysine by the addition of a solution of lOO ⁇ l/well of 50 ⁇ g/ml solution. After incubation for 1 hour at 37°C, the solution was flicked out of the wells and the plates air-dried.
  • a solution (50 ⁇ g/ml) in PBS, pH 7.4 of the polysulphated polysaccharide pentosan polysulphate or dextran sulphate was pipetted into the poly-L-lysine coated wells.
  • the saturation curve obtained using the various dilutions of MAb 5-B-10 (or other MAbs) made up in normal human serum which were incubated in wells of the pentosan polysulphate coated plates is shown in Figure 4. From this curve the dilution at 1:400 was selected to be the, most appropriate to be used in the assay.
  • Figure 5 shows the saturation curve obtained for various concentrations of pentosan polysulphate used for coating wells at 37°C for 1 hour. The concentration of 25 ⁇ g/ml of pentosan polysulphate in PBS at pH 7.4 was selected to be the most appropriate and was used thereafter in the standard assay.
  • the percentage recovery of polysulphated polysaccharides by this method was determined by adding known amounts of pentosan polysulphate to pool normal sera and assaying levels by using the standard procedure. It was found that the precentage of recovery using our method was 92.65 + 29.85%.
  • HGE human granulocyte elastase
  • BSA bovine serum albumin
  • Triton X100 0.1% (w.v.) BSA (bovine serum albumin) 0.02% (v/v) Triton X100.
  • Substrate 0.2mM Succinyl-alanine-alanine-valyl- nitroanaline (SAAVNA) added as lOO ⁇ l of 0.5mM SAAVNA in 12.5% DMSO/87.5% assay buffer. This gave a final assay concentration of 5% DMSO (dimethylsulfoxide) .
  • Inhibitor Diluted to less than lO ⁇ g/ml with assay buffer. Then assayed as serial dilutions of this concentration. Added as lOO ⁇ l in assay buffer.
  • Enzyme 0.2 ⁇ g HGE added in 50 ⁇ l assay buffer. lOO ⁇ l inhibitor and 50 ⁇ l enzyme were added to microtiter plate wells. The plate was incubated at 37°C for 5 minutes prior to initiation of the reaction by addition of prewarmed substrate solution. Absorbances at 405nm were read at regular intervals and the change in absorbance per hour calculated. The inhibitor concentration giving 50% inhibition of enzyme activity compared to an uninhibited control was calculated (IC 5Q ).
  • the monoclonal antibodies of this invention can be used in the detection and/or quantification of polysulphated polysaccharides in biological samples, in the methods of the invention and using the diagnostic kits of the invention.
  • the antibodies of the invention may be used to visualise or localise polysulphated polysaccharides in histological preparations at cell preparations.
  • the antibodies are also of use in the selection of potential drugs with anti-arthritc, anti-inflammatory, anti- degenerative, anti-coagulant and/or anti-viral activities.
  • Baba, M. , Nakajima, M. , Schols, D., Pauwels, R., Balzarini, J. and de Clercq E. Pentosan polysulfate, a sulfated aligosaccharide, is a potent and selective anti-HIV agent in vitro. Antiviral. Res. 9: 335-343, 1988b. Baba, M. Pauwels, R., Balzamini, J. , Arnaut, J. , Desmyter, J. and de Clercq, E. Mechamism of inhibitory effect of dextran sulfate and heparin on replication of human immunodeficiency virus in vitro. Proc. Nalt.
  • MacGregor I.V. Dawes J. , Paton L., Pepper D.S., Prowse C.V. and Smith M. Metabolism of sodium pentosan polysulphate in man-catabolism of iodinated derivatives. Throm Haemostas 5_L, 321-325, 1984. MacGregor, I.v., Dawes J., Paton L, Pepper D.S., Prowse C.V. and Stocks J, .

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Abstract

Ces anticorps contiennent une substitution d'anneau de disulfate-ester-pyranose 2,3, 2,6 ou 4,6. Ils sont utilisés dans les analyses de détection et/ou de quantification des polysaccharides polysulfatés contenant une substitution d'anneau de disulfate-ester-pyranose 2,3, 2,6 ou 4,6 dans les fluides biologiques. L'invention porte également sur des trousses de diagnostic servant à réaliser ces analyses.
PCT/AU1989/000547 1988-12-19 1989-12-19 Anticorps monoclonaux reconnaissant les polysaccharides polysulfates WO1990006954A1 (fr)

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0535347A2 (fr) * 1991-08-30 1993-04-07 Nalco Chemical Company Anticorps monoclonaux contre des polymères sulphonés
WO1994003530A1 (fr) * 1992-08-10 1994-02-17 Mouritsen & Elsner A/S Procede de couplage chimique sur des phases solides
EP0717995A3 (fr) * 1994-10-11 1997-04-23 Sanwa Kagaku Kenkyusho Co Agent supprimant la dégénération de cellules et reduisant la toxicité des organes
EP0869362A2 (fr) * 1997-04-04 1998-10-07 Seikagaku Corporation Procédé de détermination quantitatif d'héparansulfate et procédé diagnostic le contenant
WO1998046258A2 (fr) * 1997-04-11 1998-10-22 Beth Israel Deaconess Medical Center, Inc. Utilisation de chondroitinase dans la fabrication d'un medicament destine au traitement et a la prevention de secretions mucoides
WO2004044198A1 (fr) * 2002-11-12 2004-05-27 Taisho Pharmaceutical Co., Ltd. Anticorps reconnaissant le sulfate b de chondroitine et le sulfate e de chondroitine
WO2015152280A1 (fr) * 2014-03-31 2015-10-08 生化学工業株式会社 Anticorps anti-sulfate de chondroïtine e
US9380781B2 (en) 2010-08-26 2016-07-05 Agrosavfe N.V. Compositions for seed treatment
WO2016170158A1 (fr) * 2015-04-24 2016-10-27 Bene Pharmachem Gmbh & Co. Kg Procédé de détection et/ou de quantification de pentosane polysulfate de sodium

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4704356A (en) * 1984-03-27 1987-11-03 Rush-Presbyterian-St. Luke's Medical Center Method of diagnosing cartilage tissue abnormalities
JPS62294968A (ja) * 1986-06-16 1987-12-22 Seikagaku Kogyo Co Ltd モノクロ−ナル抗体からなる診断用薬剤
JPS63137695A (ja) * 1986-11-28 1988-06-09 Seikagaku Kogyo Co Ltd 抗コンドロイチン硫酸ｄモノクロ−ン抗体及びこれを用いるコンドロイチン硫酸ｄの検出方法
AU1625888A (en) * 1987-03-04 1988-09-26 Rockefeller University, The Monoclonal antibodies produced against glomerular proteoglycans
EP0313538A2 (fr) * 1987-10-22 1989-04-26 Carmeda Ab Anticorps monoclonal, son procédé de préparation, et son utilisation

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4704356A (en) * 1984-03-27 1987-11-03 Rush-Presbyterian-St. Luke's Medical Center Method of diagnosing cartilage tissue abnormalities
JPS62294968A (ja) * 1986-06-16 1987-12-22 Seikagaku Kogyo Co Ltd モノクロ−ナル抗体からなる診断用薬剤
JPS63137695A (ja) * 1986-11-28 1988-06-09 Seikagaku Kogyo Co Ltd 抗コンドロイチン硫酸ｄモノクロ−ン抗体及びこれを用いるコンドロイチン硫酸ｄの検出方法
AU1625888A (en) * 1987-03-04 1988-09-26 Rockefeller University, The Monoclonal antibodies produced against glomerular proteoglycans
EP0313538A2 (fr) * 1987-10-22 1989-04-26 Carmeda Ab Anticorps monoclonal, son procédé de préparation, et son utilisation

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ANALYTICAL BIOCHEMISTRY, Volume 174, 1988, pages 448-458, GOLDBERG, R.L., "Enzyme linked Immunosorbent Assay for Hyaluronate using cartilage proteoglycan and an Antibody to keratan sulfate". *
CHEMICAL ABSTRACTS, Volume 109, No. 17, issued 24 October 1988, KOIKE, Y. et al.: "Heparan sulfate determination with monoclonal antibody to heparan sulfate produced by the hybridoma method", page 385, abstract no. 145844g. *
PATENT ABSTRACTS OF JAPAN, C537, page 85; & JP,A,63 137 695, (SEIKAGAKU KOGYO CO LTD), 9 June 1988. *
PATENT ABSTRACTS OF JAPAN, P711, page 31; & JP,A,62 294 968, (SEIKAGAKU KOGYO CO LTD), 22 December 1987. *
THE JOURNAL OF BIOLOGICAL CHEMISTRY, Volume 263, Number 11, 15 April 1988, pages 5197-5201, PEJLER, G. et al.: "Monoclonal antibodies specific for oligosaccharides prepared by partial Nitrous acid Deamination of Heparin". *

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0535347A3 (en) * 1991-08-30 1993-10-27 Nalco Chemical Co Monoclonal antibodies to sulfonated polymers
EP0535347A2 (fr) * 1991-08-30 1993-04-07 Nalco Chemical Company Anticorps monoclonaux contre des polymères sulphonés
WO1994003530A1 (fr) * 1992-08-10 1994-02-17 Mouritsen & Elsner A/S Procede de couplage chimique sur des phases solides
US6262256B1 (en) 1992-08-10 2001-07-17 Mouritsen & Elsner A/S Article for tuesyl-activated dextran for solid-phase coupling
US5874569A (en) * 1992-08-10 1999-02-23 Mouritsen & Elsner A/S Method of preparing tresyl-activated dextran, article having tresyl-activated dextran fixed covalently to its surface, and immobilization of chemical compounds thereto
EP0717995A3 (fr) * 1994-10-11 1997-04-23 Sanwa Kagaku Kenkyusho Co Agent supprimant la dégénération de cellules et reduisant la toxicité des organes
US6150115A (en) * 1997-04-04 2000-11-21 Seikagaku Kogyo Kabushiki Kaisha Quantitative determination method for heparan sulfate and diagnostic method using the same
EP0869362A2 (fr) * 1997-04-04 1998-10-07 Seikagaku Corporation Procédé de détermination quantitatif d'héparansulfate et procédé diagnostic le contenant
EP0869362A3 (fr) * 1997-04-04 1998-10-14 Seikagaku Corporation Procédé de détermination quantitatif d'héparansulfate et procédé diagnostic le contenant
US6200564B1 (en) 1997-04-11 2001-03-13 J. Thomas Lamont Pharmaceutical compositions containing chondroitinase and uses therefor
WO1998046258A3 (fr) * 1997-04-11 1999-03-11 Beth Israel Hospital Utilisation de chondroitinase dans la fabrication d'un medicament destine au traitement et a la prevention de secretions mucoides
WO1998046258A2 (fr) * 1997-04-11 1998-10-22 Beth Israel Deaconess Medical Center, Inc. Utilisation de chondroitinase dans la fabrication d'un medicament destine au traitement et a la prevention de secretions mucoides
WO2004044198A1 (fr) * 2002-11-12 2004-05-27 Taisho Pharmaceutical Co., Ltd. Anticorps reconnaissant le sulfate b de chondroitine et le sulfate e de chondroitine
US9380781B2 (en) 2010-08-26 2016-07-05 Agrosavfe N.V. Compositions for seed treatment
US9516879B2 (en) 2010-08-26 2016-12-13 Agrosavfe N.V. Chitinous polysaccharide antigen-binding proteins
EP2609115B1 (fr) * 2010-08-26 2017-05-31 Agrosavfe N.V. Compositions pour le traitement de graines
WO2015152280A1 (fr) * 2014-03-31 2015-10-08 生化学工業株式会社 Anticorps anti-sulfate de chondroïtine e
US9745386B2 (en) 2014-03-31 2017-08-29 Seikagaku Corporation Anti-chondroitin-sulfate-E antibody
EP3127922A4 (fr) * 2014-03-31 2017-11-08 Seikagaku Corporation Anticorps anti-sulfate de chondroïtine e
WO2016170158A1 (fr) * 2015-04-24 2016-10-27 Bene Pharmachem Gmbh & Co. Kg Procédé de détection et/ou de quantification de pentosane polysulfate de sodium

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