Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_130662,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_150756,IEDB_150901,IEDB_151531,IEDB_151769,IEDB_156486,IEDB_164045

• Article ID: 905
  Kosma P, Reiter A, Hofinger A, Brade L, Brade H "Synthesis of neoglycoproteins containing Kdo epitopes specific for Chlamydophila psittaci lipopolysaccharide" - Journal of Endotoxin Research 6(1) (2000) 57-69
  

  Lipopolysaccharide, synthesis, Kdo, epitope, epitopes, specific, neoglycoprotein, neoglycoproteins

  Journal NLM ID: 9433350
  Publisher: Maney Publishing
  Correspondence: pkosma@edv2.boku.ac.at
  Institutions: Department of Chemistry, University of Agricultural Sciences, Muthgasse, Vienna, Austria
  Methods: NMR
  
   
  
  Chlamydophila psittaci
  CSDB ID 3003 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 1029
  Müller-Loennies S, Grimmecke D, Brade L, Lindner B, Kosma P, Brade H "A novel strategy for the synthesis of neoglycoconjugates from deacylated deep rough lipopolysaccharides" - Journal of Endotoxin Research 8(4) (2002) 295-305
  

  We report a novel strategy for the preparation of neoglycoconjugates of oligosaccharides which are obtained after complete deacylation of bacterial deep rough lipopolysaccharides (LPS) isolated from recombinant Escherichia coli bacteria synthesizing a Kdo di-[a-Kdo-(24)-a-Kdo-(2] and a Kdo trisaccharide [a-Kdo-(28)-a-Kdo-(24)-a-Kdo-(2] of Re-type and chlamydial LPS, respectively. Unlike acylated LPS, such oligosaccharides can be obtained in pure form and thus lead to well-defined neoglycoconjugates. Cleavage of the 1-phosphate of the lipid A moiety by alkaline phosphatase treatment leads to a free reducing glucosamine which can be further reacted with allylamine. After reductive amination, spacer elongation of the allyl group with cysteamine and activation with thiophosgene, the ligands were reacted with BSA. We have compared the immunological reactivity of such defined neoglycoconjugates obtained from natural sources with those obtained by chemical synthesis and report that such neoglycoconjugates are immunogenic and well suited as antigens for the study of epitope specificities of monoclonal antibodies. In addition, we have compared these conjugates with those in which ligands were coupled by glutardialdehyde to BSA. Our approach proved to be superior since the latter led upon immunization of mice to a relatively high percentage of antibodies that reacted with glutardialdehyde derivatized BSA without the carbohydrate ligand. This was not the case for cysteamine-spacered ligands coupled via their isothiocyanate-derivatives.

  Lipopolysaccharide, synthesis, lipopolysaccharides, LPS, oligosaccharide, Bacterial, trisaccharide, group, form, Escherichia, Escherichia coli, Kdo, lipid, lipid A, Oligosaccharides, bacteria, mutant, lead, alkaline, rough, treatment, deacylated, deacylation, reducing, preparation, recombinant, acylated, Glucosamine, spacer, phosphatase, free, cleavage, chlamydial, alkaline phosphatase, amination, reductive, neoglycoconjugate, allyl, deacylated LPS, elongation, neoglycocnjugate

  NCBI PubMed ID: 12230919
  Journal NLM ID: 9433350
  Publisher: Maney Publishing
  Correspondence: hbrade@fz-borstel.de
  Institutions: Divisions of 1Biochemical and Medical Microbiology and Biophysics, Center for Medicine and Biosciences, Research Center Borstel, Borstel, Germany, Institute of Chemistry, University of Agricultural Sciences, Vienna, Austria
  Methods: NMR, MS
  
   
  
  Escherichia coli
  CSDB ID 3809 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 1029
  Müller-Loennies S, Grimmecke D, Brade L, Lindner B, Kosma P, Brade H "A novel strategy for the synthesis of neoglycoconjugates from deacylated deep rough lipopolysaccharides" - Journal of Endotoxin Research 8(4) (2002) 295-305
  

  We report a novel strategy for the preparation of neoglycoconjugates of oligosaccharides which are obtained after complete deacylation of bacterial deep rough lipopolysaccharides (LPS) isolated from recombinant Escherichia coli bacteria synthesizing a Kdo di-[a-Kdo-(24)-a-Kdo-(2] and a Kdo trisaccharide [a-Kdo-(28)-a-Kdo-(24)-a-Kdo-(2] of Re-type and chlamydial LPS, respectively. Unlike acylated LPS, such oligosaccharides can be obtained in pure form and thus lead to well-defined neoglycoconjugates. Cleavage of the 1-phosphate of the lipid A moiety by alkaline phosphatase treatment leads to a free reducing glucosamine which can be further reacted with allylamine. After reductive amination, spacer elongation of the allyl group with cysteamine and activation with thiophosgene, the ligands were reacted with BSA. We have compared the immunological reactivity of such defined neoglycoconjugates obtained from natural sources with those obtained by chemical synthesis and report that such neoglycoconjugates are immunogenic and well suited as antigens for the study of epitope specificities of monoclonal antibodies. In addition, we have compared these conjugates with those in which ligands were coupled by glutardialdehyde to BSA. Our approach proved to be superior since the latter led upon immunization of mice to a relatively high percentage of antibodies that reacted with glutardialdehyde derivatized BSA without the carbohydrate ligand. This was not the case for cysteamine-spacered ligands coupled via their isothiocyanate-derivatives.

  Lipopolysaccharide, synthesis, lipopolysaccharides, LPS, oligosaccharide, Bacterial, trisaccharide, group, form, Escherichia, Escherichia coli, Kdo, lipid, lipid A, Oligosaccharides, bacteria, mutant, lead, alkaline, rough, treatment, deacylated, deacylation, reducing, preparation, recombinant, acylated, Glucosamine, spacer, phosphatase, free, cleavage, chlamydial, alkaline phosphatase, amination, reductive, neoglycoconjugate, allyl, deacylated LPS, elongation, neoglycocnjugate

  NCBI PubMed ID: 12230919
  Journal NLM ID: 9433350
  Publisher: Maney Publishing
  Correspondence: hbrade@fz-borstel.de
  Institutions: Divisions of 1Biochemical and Medical Microbiology and Biophysics, Center for Medicine and Biosciences, Research Center Borstel, Borstel, Germany, Institute of Chemistry, University of Agricultural Sciences, Vienna, Austria
  Methods: NMR, MS
  
   
  
  Escherichia coli
  CSDB ID 3810 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 1103
  Peterson EM, de la Maza L, Brade L, Brade H "Characterization of a neutralizing monoclonal antibody directed at the lipopolysaccharide of Chlamydia pneumoniae" - Infection and Immunity 66(8) (1998) 3848-3855
  

  Identification of protective epitopes is one of the first steps in the development of a subunit vaccine. One approach to accomplishing this is to identify structures or epitopes by using monoclonal antibodies (MAb) that can attenuate infectivity in vitro and in vivo. To date attempts to use this approach with Chlamydia pneumoniae have failed. This report is the first description of a MAb directed to the lipopolysaccharide (LPS) of Chlamydia that neutralizes both in vitro and in vivo the infectivity of C. pneumoniae. MAb CP-33, an immunoglobulin G2b (IgG2b), was identified from a fusion using splenocytes from mice immunized with C. pneumoniae TW-183. By Western blot analysis, MAb CP-33 exhibited genus-specific reactivity in that it recognized the LPSs of C. pneumoniae, Chlamydia trachomatis, and Chlamydia psittaci. MAb CP-33 did not react with 15 genera of gram-negative and gram-positive bacteria and Candida albicans. By using isolated LPS of Re mutants of Escherichia coli, Salmonella enterica serovar Minnesota, and recombinants expressing the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferase gene kdtA of C. trachomatis, MAb CP-33 was shown to require for binding the presence of the genus-specific trisaccharide epitope αKdo(2→8)αKdo(2→4)αKdo. By employing synthetic oligosaccharides and neoglycoconjugates in an enzyme immunoassay (EIA) and EIA inhibition, it was further shown that MAb CP-33 differed from the extensively investigated prototype chlamydial LPS MAb S25-23. Most likely, MAb CP-33 recognizes a conformational epitope in which the αKdo(2→8)αKdo(2→4)αKdo trisaccharide is an essential structural component. When tested in an in vitro neutralization assay, MAb CP-33 gave a 50% neutralization titer of 8 ng/ml against C. pneumoniae TW-183. However, this MAb did not neutralize other C. pneumoniae strains, C. trachomatis, or C. psittaci. C. pneumoniae TW-183 was treated with either MAb CP-33 or a control IgG and then used to inoculate mice by the respiratory route. Five days after inoculation, there was a difference between the mice inoculated with the control IgG-treated inoculum and those inoculated with the MAb CP-33-treated organisms as to the number of mice infected as well as the number of inclusion-forming units recovered from lung cultures (P < 0.05). In summary, a Chlamydia-specific LPS MAb was able to neutralize in vitro the infectivity of C. pneumoniae TW-183.

  Lipopolysaccharide, core, characterization, antibodies, antibody, epitope, monoclonal, monoclonal antibodies, monoclonal antibody, Chlamydia, protective, neutralizing

  NCBI PubMed ID: 9673271
  Journal NLM ID: 0246127
  Publisher: American Society for Microbiology
  Correspondence: epeterso@uci.edu
  Institutions: Department of Pathology, University of California, Irvine , Ervine, California 92697-4800, Forschungsinstitut Borstel, Zentrum fur Medizin und Biowiddenschaften, Medizinische und Biochemische Mikrobiologie, 23845 Borstel, Germany
  Methods: serological methods
  
   
  
  Chlamydia pneumoniae
  CSDB ID 4146 (all data & tools)
• Article ID: 2211
  Bock K, Thomsen JU, Kosma P, Christian R, Holst O, Brade H "A nuclear magnetic resonance spectroscopic investigation of Kdo-containing oligosaccharides related to the genus-specific epitope of Chlamydia lipopolysaccharides" - Carbohydrate Research 229 (1992) 213-224
  

  The 1H- and 13C-NMR parameters, chemical shifts and coupling constants, for the pentasaccharide of the genus-specific epitope of Chlamydia lipopolysaccharide and related di-, tri-, and tetra-saccharides have been measured and assigned completely using 1D and 2D techniques, and their structures have been confirmed. NOE experiments indicated the preferred conformation of the pentasaccharide and the component oligosaccharides. The 3JH,H demonstrate a change in conformation by rotation of the C-6-C-7 bond of the side chain of the (2→8)-linked Kdo (unit b) in α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→6)-β-GlcN-(1→6)-GlcNol, α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→6)-β-GlcNAc-(1→O)-allyl, and α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→O)-allyl relative to that preferred in α-Kdo-(2→4)-α-Kdo-(2→6)-β-GlcNAc-(1→O)-allyl, α-Kdo-(2→8)-α-Kdo-(2→O)-allyl, α-Kdo-(2→4)-α-Kdo-(2→O)-allyl, and α-Kdo-(2→6)-β-GlcNAc-(1→O)-allyl, irrespective of the size of the aglycon, e.g., allyl or β-D-GlcN residues. The conformational results have been substantiated by computer calculations using the HSEA approach.

  NCBI PubMed ID: 1382853
  Publication DOI: 10.1016/s0008-6215(00)90571-8
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Institutions: Department of Chemistry, Carlsberg Laboratory, Valby, Denmark
  Methods: 13C NMR, 1H NMR, NMR-2D
  
   
  
  Chlamydia
  CSDB ID 116692 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 1103
  Peterson EM, de la Maza L, Brade L, Brade H "Characterization of a neutralizing monoclonal antibody directed at the lipopolysaccharide of Chlamydia pneumoniae" - Infection and Immunity 66(8) (1998) 3848-3855
  

  Identification of protective epitopes is one of the first steps in the development of a subunit vaccine. One approach to accomplishing this is to identify structures or epitopes by using monoclonal antibodies (MAb) that can attenuate infectivity in vitro and in vivo. To date attempts to use this approach with Chlamydia pneumoniae have failed. This report is the first description of a MAb directed to the lipopolysaccharide (LPS) of Chlamydia that neutralizes both in vitro and in vivo the infectivity of C. pneumoniae. MAb CP-33, an immunoglobulin G2b (IgG2b), was identified from a fusion using splenocytes from mice immunized with C. pneumoniae TW-183. By Western blot analysis, MAb CP-33 exhibited genus-specific reactivity in that it recognized the LPSs of C. pneumoniae, Chlamydia trachomatis, and Chlamydia psittaci. MAb CP-33 did not react with 15 genera of gram-negative and gram-positive bacteria and Candida albicans. By using isolated LPS of Re mutants of Escherichia coli, Salmonella enterica serovar Minnesota, and recombinants expressing the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferase gene kdtA of C. trachomatis, MAb CP-33 was shown to require for binding the presence of the genus-specific trisaccharide epitope αKdo(2→8)αKdo(2→4)αKdo. By employing synthetic oligosaccharides and neoglycoconjugates in an enzyme immunoassay (EIA) and EIA inhibition, it was further shown that MAb CP-33 differed from the extensively investigated prototype chlamydial LPS MAb S25-23. Most likely, MAb CP-33 recognizes a conformational epitope in which the αKdo(2→8)αKdo(2→4)αKdo trisaccharide is an essential structural component. When tested in an in vitro neutralization assay, MAb CP-33 gave a 50% neutralization titer of 8 ng/ml against C. pneumoniae TW-183. However, this MAb did not neutralize other C. pneumoniae strains, C. trachomatis, or C. psittaci. C. pneumoniae TW-183 was treated with either MAb CP-33 or a control IgG and then used to inoculate mice by the respiratory route. Five days after inoculation, there was a difference between the mice inoculated with the control IgG-treated inoculum and those inoculated with the MAb CP-33-treated organisms as to the number of mice infected as well as the number of inclusion-forming units recovered from lung cultures (P < 0.05). In summary, a Chlamydia-specific LPS MAb was able to neutralize in vitro the infectivity of C. pneumoniae TW-183.

  Lipopolysaccharide, core, characterization, antibodies, antibody, epitope, monoclonal, monoclonal antibodies, monoclonal antibody, Chlamydia, protective, neutralizing

  NCBI PubMed ID: 9673271
  Journal NLM ID: 0246127
  Publisher: American Society for Microbiology
  Correspondence: epeterso@uci.edu
  Institutions: Department of Pathology, University of California, Irvine , Ervine, California 92697-4800, Forschungsinstitut Borstel, Zentrum fur Medizin und Biowiddenschaften, Medizinische und Biochemische Mikrobiologie, 23845 Borstel, Germany
  Methods: serological methods
  
   
  
  Chlamydia pneumoniae
  CSDB ID 4147 (all data & tools)
• Article ID: 3281
  Kosma P, Strobl M, Allmaier G, Schmidt E, Brade H "Synthesis of pentasaccharide core structure corresponding to the genus-specific lipopolysaccharide epitope of Chlamydia" - Carbohydrate Research 254 (1994) 105-132
  

  The trisaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (16a and 16b), the tetrasaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→4)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (19a and 19b), and the pentasaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→8)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→4)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (23a and 23b) were prepared. The glycosidic linkages were formed using 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose (6) and FeCl3 as promoter as well as per-O-acetylated Kdo mono- and di-saccharide bromide derivatives (12 and 20) under Helferich conditions. The oligosaccharides, which correspond to dephosphorylated part-structures of enterobacterial and chlamydial lipopolysaccharides, were characterized by NMR spectroscopy as well as plasma desorption and matrix-assisted laser desorption mass spectrometry.

  Lipopolysaccharide, NMR, synthesis, structure, core, epitope, Chlamydia, mass spectrometry

  NCBI PubMed ID: 7514094
  Publication DOI: 10.1016/0008-6215(94)84246-9
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Institutions: Institut fur Chemie der Universitat fur Bodenkultur, Wien, Austria.
  Methods: 13C NMR, 1H NMR, NMR-2D, MALDI-MS, chemical synthesis
  
   
  
  Chlamydia
  CSDB ID 22411 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135394,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_150077,IEDB_150760,IEDB_150908,IEDB_151531,IEDB_151767,IEDB_156486,IEDB_164045

• Article ID: 1103
  Peterson EM, de la Maza L, Brade L, Brade H "Characterization of a neutralizing monoclonal antibody directed at the lipopolysaccharide of Chlamydia pneumoniae" - Infection and Immunity 66(8) (1998) 3848-3855
  

  Identification of protective epitopes is one of the first steps in the development of a subunit vaccine. One approach to accomplishing this is to identify structures or epitopes by using monoclonal antibodies (MAb) that can attenuate infectivity in vitro and in vivo. To date attempts to use this approach with Chlamydia pneumoniae have failed. This report is the first description of a MAb directed to the lipopolysaccharide (LPS) of Chlamydia that neutralizes both in vitro and in vivo the infectivity of C. pneumoniae. MAb CP-33, an immunoglobulin G2b (IgG2b), was identified from a fusion using splenocytes from mice immunized with C. pneumoniae TW-183. By Western blot analysis, MAb CP-33 exhibited genus-specific reactivity in that it recognized the LPSs of C. pneumoniae, Chlamydia trachomatis, and Chlamydia psittaci. MAb CP-33 did not react with 15 genera of gram-negative and gram-positive bacteria and Candida albicans. By using isolated LPS of Re mutants of Escherichia coli, Salmonella enterica serovar Minnesota, and recombinants expressing the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferase gene kdtA of C. trachomatis, MAb CP-33 was shown to require for binding the presence of the genus-specific trisaccharide epitope αKdo(2→8)αKdo(2→4)αKdo. By employing synthetic oligosaccharides and neoglycoconjugates in an enzyme immunoassay (EIA) and EIA inhibition, it was further shown that MAb CP-33 differed from the extensively investigated prototype chlamydial LPS MAb S25-23. Most likely, MAb CP-33 recognizes a conformational epitope in which the αKdo(2→8)αKdo(2→4)αKdo trisaccharide is an essential structural component. When tested in an in vitro neutralization assay, MAb CP-33 gave a 50% neutralization titer of 8 ng/ml against C. pneumoniae TW-183. However, this MAb did not neutralize other C. pneumoniae strains, C. trachomatis, or C. psittaci. C. pneumoniae TW-183 was treated with either MAb CP-33 or a control IgG and then used to inoculate mice by the respiratory route. Five days after inoculation, there was a difference between the mice inoculated with the control IgG-treated inoculum and those inoculated with the MAb CP-33-treated organisms as to the number of mice infected as well as the number of inclusion-forming units recovered from lung cultures (P < 0.05). In summary, a Chlamydia-specific LPS MAb was able to neutralize in vitro the infectivity of C. pneumoniae TW-183.

  Lipopolysaccharide, core, characterization, antibodies, antibody, epitope, monoclonal, monoclonal antibodies, monoclonal antibody, Chlamydia, protective, neutralizing

  NCBI PubMed ID: 9673271
  Journal NLM ID: 0246127
  Publisher: American Society for Microbiology
  Correspondence: epeterso@uci.edu
  Institutions: Department of Pathology, University of California, Irvine , Ervine, California 92697-4800, Forschungsinstitut Borstel, Zentrum fur Medizin und Biowiddenschaften, Medizinische und Biochemische Mikrobiologie, 23845 Borstel, Germany
  Methods: serological methods
  
   
  
  Chlamydia pneumoniae F515-207
  CSDB ID 4148 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 2592
  Holst O, Brade L, Kosma P, Brade H "Structure, serological specificity, and synthesis of artificial glycoconjugates representing the genus-specific lipopolysaccharide epitope of Chlamydia spp" - Journal of Bacteriology 173 (1991) 1862-1866
  
  Journal NLM ID: 2985120R
  Publisher: American Society for Microbiology
  
   
  
  Chlamydia
  CSDB ID 131773 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: BSA
Trivial name: neoglycoconjugate
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_130662,IEDB_135394,IEDB_135813,IEDB_137340,IEDB_140092,IEDB_141807,IEDB_150077,IEDB_150755,IEDB_150756,IEDB_150760,IEDB_150901,IEDB_150908,IEDB_151531,IEDB_151767,IEDB_151768,IEDB_151769,IEDB_156486,IEDB_156487,IEDB_164045

• Article ID: 3163
  Müller-Loennies S, Gronow S, Brade L, Mackenzie R, Kosma P, Brade H "A monoclonal antibody against a carbohydrate epitope in lipopolysaccharide differentiates Chlamydophila psittaci from Chlamydophila pecorum, Chlamydophila pneumoniae, and Chlamydia trachomatis" - Glycobiology 16(3) (2006) 184-196
  

  Lipopolysaccharide (LPS) of Chlamydophila psittaci but not of Chlamydophila pneumoniae or Chlamydia trachomatis contains a tetrasaccharide of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) of the sequence Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo. After immunization with the synthetic neoglycoconjugate antigen Kdo(2→8)[Kdo(2→4)]Kdo(2→4) Kdo-BSA, we obtained the mouse monoclonal antibody (mAb) S69-4 which was able to differentiate C. psittaci from Chlamydophila pecorum, C. pneumoniae, and C. trachomatis in double labeling experiments of infected cell monolayers and by enzyme-linked immunosorbent assay (ELISA). The epitope specificity of mAb S69-4 was determined by binding and inhibition assays using bacteria, LPS, and natural or synthetic Kdo oligosaccharides as free ligands or conjugated to BSA. The mAb bound preferentially Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo(2→4) with a K(d) of 10 microM, as determined by surface plasmon resonance (SPR) for the monovalent interaction using mAb or single chain Fv. Cross-reactivity was observed with Kdo(2→4)Kdo(2→4)Kdo but not with Kdo(2→8)Kdo(2→4)Kdo, Kdo disaccharides in 2→4- or 2→8-linkage, or Kdo monosaccharide. MAb S69-4 was able to detect LPS on thin-layer chromatography (TLC) plates in amounts of <10 ng by immunostaining. Due to the high sensitivity achieved in this assay, the antibody also detected in vitro products of cloned Kdo transferases of Chlamydia. The antibody can therefore be used in medical and veterinarian diagnostics, general microbiology, analytical biochemistry, and studies of chlamydial LPS biosynthesis. Further contribution to the general understanding of carbohydrate-binding antibodies was obtained by a comparison of the primary structure of mAb S69-4 to that of mAb S45-18 of which the crystal structure in complex with its ligand has been elucidated recently (Nguyen et al., 2003, Nat. Struct. Biol., 10, 1019-1025).

  Kdo, diagnostic, neoglycoconjugate, immunofluorescence

  NCBI PubMed ID: 16282606
  Publication DOI: 10.1093/glycob/cwj055
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany, Institute for Biological Sci-ences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austri
  Methods: serological methods, genetic methods
  
   
  
  Escherichia coli, Chlamydophila psittaci
  CSDB ID 20498 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: BSA
Trivial name: neoglycoconjugate
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_151767,IEDB_156486,IEDB_164045

• Article ID: 3163
  Müller-Loennies S, Gronow S, Brade L, Mackenzie R, Kosma P, Brade H "A monoclonal antibody against a carbohydrate epitope in lipopolysaccharide differentiates Chlamydophila psittaci from Chlamydophila pecorum, Chlamydophila pneumoniae, and Chlamydia trachomatis" - Glycobiology 16(3) (2006) 184-196
  

  Lipopolysaccharide (LPS) of Chlamydophila psittaci but not of Chlamydophila pneumoniae or Chlamydia trachomatis contains a tetrasaccharide of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) of the sequence Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo. After immunization with the synthetic neoglycoconjugate antigen Kdo(2→8)[Kdo(2→4)]Kdo(2→4) Kdo-BSA, we obtained the mouse monoclonal antibody (mAb) S69-4 which was able to differentiate C. psittaci from Chlamydophila pecorum, C. pneumoniae, and C. trachomatis in double labeling experiments of infected cell monolayers and by enzyme-linked immunosorbent assay (ELISA). The epitope specificity of mAb S69-4 was determined by binding and inhibition assays using bacteria, LPS, and natural or synthetic Kdo oligosaccharides as free ligands or conjugated to BSA. The mAb bound preferentially Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo(2→4) with a K(d) of 10 microM, as determined by surface plasmon resonance (SPR) for the monovalent interaction using mAb or single chain Fv. Cross-reactivity was observed with Kdo(2→4)Kdo(2→4)Kdo but not with Kdo(2→8)Kdo(2→4)Kdo, Kdo disaccharides in 2→4- or 2→8-linkage, or Kdo monosaccharide. MAb S69-4 was able to detect LPS on thin-layer chromatography (TLC) plates in amounts of <10 ng by immunostaining. Due to the high sensitivity achieved in this assay, the antibody also detected in vitro products of cloned Kdo transferases of Chlamydia. The antibody can therefore be used in medical and veterinarian diagnostics, general microbiology, analytical biochemistry, and studies of chlamydial LPS biosynthesis. Further contribution to the general understanding of carbohydrate-binding antibodies was obtained by a comparison of the primary structure of mAb S69-4 to that of mAb S45-18 of which the crystal structure in complex with its ligand has been elucidated recently (Nguyen et al., 2003, Nat. Struct. Biol., 10, 1019-1025).

  Kdo, diagnostic, neoglycoconjugate, immunofluorescence

  NCBI PubMed ID: 16282606
  Publication DOI: 10.1093/glycob/cwj055
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany, Institute for Biological Sci-ences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austri
  Methods: serological methods, genetic methods
  
   
  
  Escherichia coli, Chlamydophila psittaci
  CSDB ID 20500 (all data & tools)

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Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_156486,IEDB_164045

• Article ID: 3281
  Kosma P, Strobl M, Allmaier G, Schmidt E, Brade H "Synthesis of pentasaccharide core structure corresponding to the genus-specific lipopolysaccharide epitope of Chlamydia" - Carbohydrate Research 254 (1994) 105-132
  

  The trisaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (16a and 16b), the tetrasaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→4)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (19a and 19b), and the pentasaccharides allyl O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→8)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→4)-O-(sodium 3-deoxy-α-D-manno-2-octulopyranosylonate)-(2→6)-O-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1→6)-2-acetamido-2-deoxy-α- and -β-D-glucopyranoside (23a and 23b) were prepared. The glycosidic linkages were formed using 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose (6) and FeCl3 as promoter as well as per-O-acetylated Kdo mono- and di-saccharide bromide derivatives (12 and 20) under Helferich conditions. The oligosaccharides, which correspond to dephosphorylated part-structures of enterobacterial and chlamydial lipopolysaccharides, were characterized by NMR spectroscopy as well as plasma desorption and matrix-assisted laser desorption mass spectrometry.

  Lipopolysaccharide, NMR, synthesis, structure, core, epitope, Chlamydia, mass spectrometry

  NCBI PubMed ID: 7514094
  Publication DOI: 10.1016/0008-6215(94)84246-9
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Institutions: Institut fur Chemie der Universitat fur Bodenkultur, Wien, Austria.
  Methods: 13C NMR, 1H NMR, NMR-2D, MALDI-MS, chemical synthesis
  
   
  
  Chlamydia
  CSDB ID 22509 (all data & tools)