Chemical conjugation of a novel antibody-interleukin 2 immunoconjugate against c-erbB-2 product

To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL-2) conjugate. MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated...

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Veröffentlicht in:	Chinese medical journal 2000-02, Vol.113 (2), p.151-153
Hauptverfasser:	Li, J, Lee, S E, Belciug, M, Ring, D B, Kwok, C S
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Antibodies, Monoclonal - chemistry Cell Division - drug effects Cell Division - immunology Cell Line Cells, Cultured Electrophoresis, Polyacrylamide Gel Humans Immunoconjugates - isolation & purification Immunoconjugates - metabolism Immunoconjugates - pharmacology Interleukin-2 - chemistry Protein Binding Receptor, ErbB-2 - immunology Recombinant Proteins - chemistry Tumor Cells, Cultured
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container_title	Chinese medical journal
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creator	Li, J Lee, S E Belciug, M Ring, D B Kwok, C S
description	To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL-2) conjugate. MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated by Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, a heterobifunctional linker, and N-succinimidyl s-acetylthioacetate was introduced onto 520C9. Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL-2 to generate 520C9-rhIL-2 immunoconjugate. The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay. The immunoconjugate also possessed IL-2 activity as measured by the standard CTLL-2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine-activated killer cells. Our method of conjugation of rhIL-2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL-2. This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines.
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MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated by Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, a heterobifunctional linker, and N-succinimidyl s-acetylthioacetate was introduced onto 520C9. Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL-2 to generate 520C9-rhIL-2 immunoconjugate. The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay. The immunoconjugate also possessed IL-2 activity as measured by the standard CTLL-2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine-activated killer cells. Our method of conjugation of rhIL-2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL-2. This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines.</description><identifier>ISSN: 0366-6999</identifier><identifier>PMID: 11775541</identifier><language>eng</language><publisher>China</publisher><subject>Animals ; Antibodies, Monoclonal - chemistry ; Cell Division - drug effects ; Cell Division - immunology ; Cell Line ; Cells, Cultured ; Electrophoresis, Polyacrylamide Gel ; Humans ; Immunoconjugates - isolation & purification ; Immunoconjugates - metabolism ; Immunoconjugates - pharmacology ; Interleukin-2 - chemistry ; Protein Binding ; Receptor, ErbB-2 - immunology ; Recombinant Proteins - chemistry ; Tumor Cells, Cultured</subject><ispartof>Chinese medical journal, 2000-02, Vol.113 (2), p.151-153</ispartof><rights>Copyright © Wanfang Data Co. Ltd. All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.wanfangdata.com.cn/images/PeriodicalImages/zhcmj/zhcmj.jpg</thumbnail><link.rule.ids>314,776,780</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11775541$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, J</creatorcontrib><creatorcontrib>Lee, S E</creatorcontrib><creatorcontrib>Belciug, M</creatorcontrib><creatorcontrib>Ring, D B</creatorcontrib><creatorcontrib>Kwok, C S</creatorcontrib><title>Chemical conjugation of a novel antibody-interleukin 2 immunoconjugate against c-erbB-2 product</title><title>Chinese medical journal</title><addtitle>Chin Med J (Engl)</addtitle><description>To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL-2) conjugate. MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated by Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, a heterobifunctional linker, and N-succinimidyl s-acetylthioacetate was introduced onto 520C9. Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL-2 to generate 520C9-rhIL-2 immunoconjugate. The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay. The immunoconjugate also possessed IL-2 activity as measured by the standard CTLL-2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine-activated killer cells. Our method of conjugation of rhIL-2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL-2. This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - chemistry</subject><subject>Cell Division - drug effects</subject><subject>Cell Division - immunology</subject><subject>Cell Line</subject><subject>Cells, Cultured</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Humans</subject><subject>Immunoconjugates - isolation & purification</subject><subject>Immunoconjugates - metabolism</subject><subject>Immunoconjugates - pharmacology</subject><subject>Interleukin-2 - chemistry</subject><subject>Protein Binding</subject><subject>Receptor, ErbB-2 - immunology</subject><subject>Recombinant Proteins - chemistry</subject><subject>Tumor Cells, Cultured</subject><issn>0366-6999</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkMtOwzAURL0A0VL4BeQVu0iOX0mWUPGSKrGBtXXj2K1DbJc4BpWvJxKtmM1szsy9mjO0JEzKQjZNs0CXKfWEUCEqeYEWZVlVQvByidR6Z7zTMGAdQ5-3MLkYcLQYcIhfZsAQJtfG7lC4MJlxMPnDBUyx8z6HeMoYDFtwIU1YF2Zs7wuK92Pssp6u0LmFIZnro6_Q--PD2_q52Lw-vazvNsWesnoqKiC1ZmVd2lrzjlvOiKgtdI0kYGRrwWhOhGHUWg6ikZVuO6EZEKtFW1nKVuj2r_cbgoWwVX3MY5gvqp-d9j0lsygp-T84P_iZTZqUd0mbYYBgYk6qIqKUnMsZvDmCufWmU_vReRgP6rQd-wWMfmrR</recordid><startdate>20000201</startdate><enddate>20000201</enddate><creator>Li, J</creator><creator>Lee, S E</creator><creator>Belciug, M</creator><creator>Ring, D B</creator><creator>Kwok, C S</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>2B.</scope><scope>4A8</scope><scope>92I</scope><scope>93N</scope><scope>PSX</scope><scope>TCJ</scope></search><sort><creationdate>20000201</creationdate><title>Chemical conjugation of a novel antibody-interleukin 2 immunoconjugate against c-erbB-2 product</title><author>Li, J ; Lee, S E ; Belciug, M ; Ring, D B ; Kwok, C S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p238t-7a08c3181f8c4d4f43058fad960ae6bfaec405e32ff4a5967cbd5c3a0fc5b7f23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - chemistry</topic><topic>Cell Division - drug effects</topic><topic>Cell Division - immunology</topic><topic>Cell Line</topic><topic>Cells, Cultured</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Humans</topic><topic>Immunoconjugates - isolation & purification</topic><topic>Immunoconjugates - metabolism</topic><topic>Immunoconjugates - pharmacology</topic><topic>Interleukin-2 - chemistry</topic><topic>Protein Binding</topic><topic>Receptor, ErbB-2 - immunology</topic><topic>Recombinant Proteins - chemistry</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, J</creatorcontrib><creatorcontrib>Lee, S E</creatorcontrib><creatorcontrib>Belciug, M</creatorcontrib><creatorcontrib>Ring, D B</creatorcontrib><creatorcontrib>Kwok, C S</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><collection>Wanfang Data Journals - Hong Kong</collection><collection>WANFANG Data Centre</collection><collection>Wanfang Data Journals</collection><collection>万方数据期刊 - 香港版</collection><collection>China Online Journals (COJ)</collection><collection>China Online Journals (COJ)</collection><jtitle>Chinese medical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, J</au><au>Lee, S E</au><au>Belciug, M</au><au>Ring, D B</au><au>Kwok, C S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Chemical conjugation of a novel antibody-interleukin 2 immunoconjugate against c-erbB-2 product</atitle><jtitle>Chinese medical journal</jtitle><addtitle>Chin Med J (Engl)</addtitle><date>2000-02-01</date><risdate>2000</risdate><volume>113</volume><issue>2</issue><spage>151</spage><epage>153</epage><pages>151-153</pages><issn>0366-6999</issn><abstract>To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL-2) conjugate. MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated by Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, a heterobifunctional linker, and N-succinimidyl s-acetylthioacetate was introduced onto 520C9. Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL-2 to generate 520C9-rhIL-2 immunoconjugate. The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay. The immunoconjugate also possessed IL-2 activity as measured by the standard CTLL-2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine-activated killer cells. Our method of conjugation of rhIL-2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL-2. This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines.</abstract><cop>China</cop><pmid>11775541</pmid><tpages>3</tpages></addata></record>
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subjects	Animals Antibodies, Monoclonal - chemistry Cell Division - drug effects Cell Division - immunology Cell Line Cells, Cultured Electrophoresis, Polyacrylamide Gel Humans Immunoconjugates - isolation & purification Immunoconjugates - metabolism Immunoconjugates - pharmacology Interleukin-2 - chemistry Protein Binding Receptor, ErbB-2 - immunology Recombinant Proteins - chemistry Tumor Cells, Cultured
title	Chemical conjugation of a novel antibody-interleukin 2 immunoconjugate against c-erbB-2 product
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