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Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide–P64k protein conjugates
Neisseria meningitidis serogroup C polysaccharide (CCPS) was conjugated to the carrier protein P64k using two different conjugation procedures, condensation mediated by carbodiimide with adipic acid dihydrazide as spacer and the reductive amination method. BALB/c mice were immunized with the resulta...
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| Published in: | FEMS immunology and medical microbiology 2004-04, Vol.40 (3), p.193-199 |
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| Main Authors: | , , , , , , , |
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| Language: | English |
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| container_end_page | 199 |
| container_issue | 3 |
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| container_title | FEMS immunology and medical microbiology |
| container_volume | 40 |
| creator | Carmenate, Tania Canaán, Leonardo Álvarez, Anabel Delgado, Maité González, Sonia Menéndez, Tamara Rodés, Lorenzo Guillén, Gerardo |
| description | Neisseria meningitidis serogroup C polysaccharide (CCPS) was conjugated to the carrier protein P64k using two different conjugation procedures, condensation mediated by carbodiimide with adipic acid dihydrazide as spacer and the reductive amination method. BALB/c mice were immunized with the resultant polysaccharide–protein conjugates and the immune response was evaluated. All conjugates assayed generated at least 10-fold higher antibody titers than the free polysaccharide. The reductive amination method rendered the best conjugate (CCPS–P64k
R) that was able to elicit antibody titers statistically higher than the titer elicited by the plain CCPS (
P |
| doi_str_mv | 10.1016/S0928-8244(03)00346-8 |
| format | article |
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R) that was able to elicit antibody titers statistically higher than the titer elicited by the plain CCPS (
P<0.001). The sera of the group immunized with CCPS–P64k
R showed a three-fold higher bactericidal response than the sera of the group immunized with the plain CCPS and they were able to protect against challenge with meningococci in the infant rat protection model. In addition, three different conjugates were obtained from polysaccharides with molecular relative sizes of 2000–4000 Da, 4000–10 000 Da or 10 000–50 000 Da, but no differences were detected in the immune response obtained against the three conjugates. Our experiments demonstrate that it is possible to generate a protective, T-cell-dependent response against CCPS using the P64k protein as carrier.</description><identifier>ISSN: 0928-8244</identifier><identifier>EISSN: 1574-695X</identifier><identifier>DOI: 10.1016/S0928-8244(03)00346-8</identifier><identifier>PMID: 15039094</identifier><language>eng</language><publisher>Oxford, UK: Elsevier B.V</publisher><subject>Adipates ; Amination ; Animals ; Antibodies, Bacterial - blood ; Bacterial Outer Membrane Proteins - chemistry ; Bacterial Outer Membrane Proteins - immunology ; Bacteriology ; Biological and medical sciences ; Carbodiimides ; Colony Count, Microbial ; Disease Models, Animal ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Immunoglobulin G - blood ; Meningococcal Infections - immunology ; Meningococcal Infections - prevention & control ; Meningococcal vaccine ; Meningococcal Vaccines - chemistry ; Meningococcal Vaccines - immunology ; Mice ; Mice, Inbred BALB C ; Microbiology ; Miscellaneous ; Molecular Weight ; Neisseria meningitidis ; Neisseria meningitidis, Serogroup C - growth & development ; Neisseria meningitidis, Serogroup C - immunology ; Polysaccharide conjugate ; Polysaccharides, Bacterial - chemistry ; Polysaccharides, Bacterial - immunology ; Rats ; Tissues, organs and organisms biophysics ; Vaccines, Conjugate - chemistry ; Vaccines, Conjugate - immunology</subject><ispartof>FEMS immunology and medical microbiology, 2004-04, Vol.40 (3), p.193-199</ispartof><rights>2003</rights><rights>2003 Federation of European Microbiological Societies. 2003</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15603682$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15039094$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carmenate, Tania</creatorcontrib><creatorcontrib>Canaán, Leonardo</creatorcontrib><creatorcontrib>Álvarez, Anabel</creatorcontrib><creatorcontrib>Delgado, Maité</creatorcontrib><creatorcontrib>González, Sonia</creatorcontrib><creatorcontrib>Menéndez, Tamara</creatorcontrib><creatorcontrib>Rodés, Lorenzo</creatorcontrib><creatorcontrib>Guillén, Gerardo</creatorcontrib><title>Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide–P64k protein conjugates</title><title>FEMS immunology and medical microbiology</title><addtitle>FEMS Immunol Med Microbiol</addtitle><description>Neisseria meningitidis serogroup C polysaccharide (CCPS) was conjugated to the carrier protein P64k using two different conjugation procedures, condensation mediated by carbodiimide with adipic acid dihydrazide as spacer and the reductive amination method. BALB/c mice were immunized with the resultant polysaccharide–protein conjugates and the immune response was evaluated. All conjugates assayed generated at least 10-fold higher antibody titers than the free polysaccharide. The reductive amination method rendered the best conjugate (CCPS–P64k
R) that was able to elicit antibody titers statistically higher than the titer elicited by the plain CCPS (
P<0.001). The sera of the group immunized with CCPS–P64k
R showed a three-fold higher bactericidal response than the sera of the group immunized with the plain CCPS and they were able to protect against challenge with meningococci in the infant rat protection model. In addition, three different conjugates were obtained from polysaccharides with molecular relative sizes of 2000–4000 Da, 4000–10 000 Da or 10 000–50 000 Da, but no differences were detected in the immune response obtained against the three conjugates. Our experiments demonstrate that it is possible to generate a protective, T-cell-dependent response against CCPS using the P64k protein as carrier.</description><subject>Adipates</subject><subject>Amination</subject><subject>Animals</subject><subject>Antibodies, Bacterial - blood</subject><subject>Bacterial Outer Membrane Proteins - chemistry</subject><subject>Bacterial Outer Membrane Proteins - immunology</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Carbodiimides</subject><subject>Colony Count, Microbial</subject><subject>Disease Models, Animal</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Immunoglobulin G - blood</subject><subject>Meningococcal Infections - immunology</subject><subject>Meningococcal Infections - prevention & control</subject><subject>Meningococcal vaccine</subject><subject>Meningococcal Vaccines - chemistry</subject><subject>Meningococcal Vaccines - immunology</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Molecular Weight</subject><subject>Neisseria meningitidis</subject><subject>Neisseria meningitidis, Serogroup C - growth & development</subject><subject>Neisseria meningitidis, Serogroup C - immunology</subject><subject>Polysaccharide conjugate</subject><subject>Polysaccharides, Bacterial - chemistry</subject><subject>Polysaccharides, Bacterial - immunology</subject><subject>Rats</subject><subject>Tissues, organs and organisms biophysics</subject><subject>Vaccines, Conjugate - chemistry</subject><subject>Vaccines, Conjugate - immunology</subject><issn>0928-8244</issn><issn>1574-695X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><recordid>eNqFkd1qFDEYhoModlu9BCUnSj0Yzf9MjkSWVgsVBRU8C9nMl93Umcl2MlOZM29BvEOvxGxnrQqiRyHkeb58vC9CDyh5SglVz94RzaqiYkIcE_6EEC5UUd1CCypLUSgtP95GixvkAB2mdEEIEZqQu-iASsI10WKBvp54D27A0WMXu4txbYcQO9zCsIl1bOJ6wvk6bACHth27uIYuuDBM2HY13vZxyHK4AgzeB2fdtBvUZqZbRxedsw1e93Hc4iXexmZK1rmN7UMN3798e6vEp3lE6G4-h3QP3fG2SXB_fx6hD6cn75evivM3L8-WL84LEFTywpes5rJ0ljDHay5WlfeMrKSUhDFngZeSAdfSKVYr6TxbVVQ7Qb2mmnmu-BF6PM_NK1yOkAbThuSgaWwHcUympKVUVPwfpKXWilKdwYd7cFy1UJttH1rbT-Zn2hl4tAdsytH43nYupN84RbiqWOb0zH0ODUy_3onZVW-uqze7Xg3h5rp6U5nTs9d5h-yS2c2p_90s_jCLKivPZwVy3FcBepNcgM5BHfpcr6lj-Pff_AcnvcVt</recordid><startdate>20040409</startdate><enddate>20040409</enddate><creator>Carmenate, Tania</creator><creator>Canaán, Leonardo</creator><creator>Álvarez, Anabel</creator><creator>Delgado, Maité</creator><creator>González, Sonia</creator><creator>Menéndez, Tamara</creator><creator>Rodés, Lorenzo</creator><creator>Guillén, Gerardo</creator><general>Elsevier B.V</general><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QL</scope><scope>7T5</scope><scope>C1K</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20040409</creationdate><title>Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide–P64k protein conjugates</title><author>Carmenate, Tania ; Canaán, Leonardo ; Álvarez, Anabel ; Delgado, Maité ; González, Sonia ; Menéndez, Tamara ; Rodés, Lorenzo ; Guillén, Gerardo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e4153-f72d357ca02c3d34b8ff20b555022cae3752e395c62d65cf2b819c41f9192f363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Adipates</topic><topic>Amination</topic><topic>Animals</topic><topic>Antibodies, Bacterial - blood</topic><topic>Bacterial Outer Membrane Proteins - chemistry</topic><topic>Bacterial Outer Membrane Proteins - immunology</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Carbodiimides</topic><topic>Colony Count, Microbial</topic><topic>Disease Models, Animal</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Immunoglobulin G - blood</topic><topic>Meningococcal Infections - immunology</topic><topic>Meningococcal Infections - prevention & control</topic><topic>Meningococcal vaccine</topic><topic>Meningococcal Vaccines - chemistry</topic><topic>Meningococcal Vaccines - immunology</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Molecular Weight</topic><topic>Neisseria meningitidis</topic><topic>Neisseria meningitidis, Serogroup C - growth & development</topic><topic>Neisseria meningitidis, Serogroup C - immunology</topic><topic>Polysaccharide conjugate</topic><topic>Polysaccharides, Bacterial - chemistry</topic><topic>Polysaccharides, Bacterial - immunology</topic><topic>Rats</topic><topic>Tissues, organs and organisms biophysics</topic><topic>Vaccines, Conjugate - chemistry</topic><topic>Vaccines, Conjugate - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Carmenate, Tania</creatorcontrib><creatorcontrib>Canaán, Leonardo</creatorcontrib><creatorcontrib>Álvarez, Anabel</creatorcontrib><creatorcontrib>Delgado, Maité</creatorcontrib><creatorcontrib>González, Sonia</creatorcontrib><creatorcontrib>Menéndez, Tamara</creatorcontrib><creatorcontrib>Rodés, Lorenzo</creatorcontrib><creatorcontrib>Guillén, Gerardo</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Immunology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>FEMS immunology and medical microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Carmenate, Tania</au><au>Canaán, Leonardo</au><au>Álvarez, Anabel</au><au>Delgado, Maité</au><au>González, Sonia</au><au>Menéndez, Tamara</au><au>Rodés, Lorenzo</au><au>Guillén, Gerardo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide–P64k protein conjugates</atitle><jtitle>FEMS immunology and medical microbiology</jtitle><addtitle>FEMS Immunol Med Microbiol</addtitle><date>2004-04-09</date><risdate>2004</risdate><volume>40</volume><issue>3</issue><spage>193</spage><epage>199</epage><pages>193-199</pages><issn>0928-8244</issn><eissn>1574-695X</eissn><abstract>Neisseria meningitidis serogroup C polysaccharide (CCPS) was conjugated to the carrier protein P64k using two different conjugation procedures, condensation mediated by carbodiimide with adipic acid dihydrazide as spacer and the reductive amination method. BALB/c mice were immunized with the resultant polysaccharide–protein conjugates and the immune response was evaluated. All conjugates assayed generated at least 10-fold higher antibody titers than the free polysaccharide. The reductive amination method rendered the best conjugate (CCPS–P64k
R) that was able to elicit antibody titers statistically higher than the titer elicited by the plain CCPS (
P<0.001). The sera of the group immunized with CCPS–P64k
R showed a three-fold higher bactericidal response than the sera of the group immunized with the plain CCPS and they were able to protect against challenge with meningococci in the infant rat protection model. In addition, three different conjugates were obtained from polysaccharides with molecular relative sizes of 2000–4000 Da, 4000–10 000 Da or 10 000–50 000 Da, but no differences were detected in the immune response obtained against the three conjugates. Our experiments demonstrate that it is possible to generate a protective, T-cell-dependent response against CCPS using the P64k protein as carrier.</abstract><cop>Oxford, UK</cop><pub>Elsevier B.V</pub><pmid>15039094</pmid><doi>10.1016/S0928-8244(03)00346-8</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0928-8244 |
| ispartof | FEMS immunology and medical microbiology, 2004-04, Vol.40 (3), p.193-199 |
| issn | 0928-8244 1574-695X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_71756146 |
| source | Oxford Journals Online |
| subjects | Adipates Amination Animals Antibodies, Bacterial - blood Bacterial Outer Membrane Proteins - chemistry Bacterial Outer Membrane Proteins - immunology Bacteriology Biological and medical sciences Carbodiimides Colony Count, Microbial Disease Models, Animal Fundamental and applied biological sciences. Psychology Fundamental immunology Immunoglobulin G - blood Meningococcal Infections - immunology Meningococcal Infections - prevention & control Meningococcal vaccine Meningococcal Vaccines - chemistry Meningococcal Vaccines - immunology Mice Mice, Inbred BALB C Microbiology Miscellaneous Molecular Weight Neisseria meningitidis Neisseria meningitidis, Serogroup C - growth & development Neisseria meningitidis, Serogroup C - immunology Polysaccharide conjugate Polysaccharides, Bacterial - chemistry Polysaccharides, Bacterial - immunology Rats Tissues, organs and organisms biophysics Vaccines, Conjugate - chemistry Vaccines, Conjugate - immunology |
| title | Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide–P64k protein conjugates |
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