The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs)
Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion protei...
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| Veröffentlicht in: | Journal of Atherosclerosis and Thrombosis 2002, Vol.9(5), pp.233-242 |
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| container_title | Journal of Atherosclerosis and Thrombosis |
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| creator | Tanaka, Toshiya Takeno, Tetsu Watanabe, Yuichiro Uchiyama, Yasutoshi Murakami, Takeshi Yamashita, Hisahiko Suzuki, Akifumi Aoi, Rie Iwanari, Hiroko Jiang, Shu-Ying Naito, Makoto Tachibana, Keisuke Doi, Takefumi Shulman, Andrew I. Mangelsdorf, David J. Reiter, Raphael Auwerx, Johan Hamakubo, Takao Kodama, Tatsuhiko |
| description | Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step. |
| doi_str_mv | 10.5551/jat.9.233 |
| format | Article |
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In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.</description><identifier>ISSN: 1340-3478</identifier><identifier>EISSN: 1880-3873</identifier><identifier>DOI: 10.5551/jat.9.233</identifier><identifier>PMID: 12409633</identifier><language>eng</language><publisher>Japan: Japan Atherosclerosis Society</publisher><subject>Animals ; Antibodies, Monoclonal - genetics ; Antibodies, Monoclonal - immunology ; Baculovirus ; Blotting, Western ; CHO Cells ; Cricetinae ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Female ; Gp64-fusion protein ; Humans ; Immunohistochemistry ; Life Sciences ; Mice ; Mice, Inbred BALB C ; Monoclonal antibody (Mab) ; Nucleopolyhedrovirus - genetics ; Peroxisome proliferator-activated receptors (PPARs) ; Receptors, Cytoplasmic and Nuclear - immunology ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - immunology ; Spodoptera ; Transcription Factors - immunology</subject><ispartof>Journal of Atherosclerosis and Thrombosis, 2002, Vol.9(5), pp.233-242</ispartof><rights>2002 Japan Atherosclerosis Society</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5103-42c42bc57d47447f9a3b790598c7f6518397b79d500e8290c051fdb6f64b5c843</citedby><cites>FETCH-LOGICAL-c5103-42c42bc57d47447f9a3b790598c7f6518397b79d500e8290c051fdb6f64b5c843</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,1883,4024,27923,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12409633$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-04151318$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Tanaka, Toshiya</creatorcontrib><creatorcontrib>Takeno, Tetsu</creatorcontrib><creatorcontrib>Watanabe, Yuichiro</creatorcontrib><creatorcontrib>Uchiyama, Yasutoshi</creatorcontrib><creatorcontrib>Murakami, Takeshi</creatorcontrib><creatorcontrib>Yamashita, Hisahiko</creatorcontrib><creatorcontrib>Suzuki, Akifumi</creatorcontrib><creatorcontrib>Aoi, Rie</creatorcontrib><creatorcontrib>Iwanari, Hiroko</creatorcontrib><creatorcontrib>Jiang, Shu-Ying</creatorcontrib><creatorcontrib>Naito, Makoto</creatorcontrib><creatorcontrib>Tachibana, Keisuke</creatorcontrib><creatorcontrib>Doi, Takefumi</creatorcontrib><creatorcontrib>Shulman, Andrew I.</creatorcontrib><creatorcontrib>Mangelsdorf, David J.</creatorcontrib><creatorcontrib>Reiter, Raphael</creatorcontrib><creatorcontrib>Auwerx, Johan</creatorcontrib><creatorcontrib>Hamakubo, Takao</creatorcontrib><creatorcontrib>Kodama, Tatsuhiko</creatorcontrib><title>The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs)</title><title>Journal of Atherosclerosis and Thrombosis</title><addtitle>JAT</addtitle><description>Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - genetics</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Baculovirus</subject><subject>Blotting, Western</subject><subject>CHO Cells</subject><subject>Cricetinae</subject><subject>Electrophoretic Mobility Shift Assay</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Female</subject><subject>Gp64-fusion protein</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Life Sciences</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Monoclonal antibody (Mab)</subject><subject>Nucleopolyhedrovirus - genetics</subject><subject>Peroxisome proliferator-activated receptors (PPARs)</subject><subject>Receptors, Cytoplasmic and Nuclear - immunology</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - immunology</subject><subject>Spodoptera</subject><subject>Transcription Factors - immunology</subject><issn>1340-3478</issn><issn>1880-3873</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkc1uGyEURlHVqvlpF32BilXVLMaFAQZm0YUVpXElR7GidI0Y5k6MNQMu4Kh5--LYchZXXH069yz4EPpCyUwIQX9sTJ61s5qxd-icKkUqpiR7X3bGy86lOkMXKW0IYUyI-iM6ozUnbcPYOXKPa8C34CGa7ILHYcB3wQc7Bm9GPPfZdaF3kLB5Ms6njBe7yXi8ghj-uRQmwKsYRjfs70OsjM3u2WTo8QNY2JYo4e-r1fwhXX1CHwYzJvh8fC_Rn183j9eLanl_-_t6vqysoIRVvLa87qyQPZecy6E1rJMtEa2ycmgEVayVJegFIaDqllgi6NB3zdDwTljF2SW6OnjXZtTb6CYTX3QwTi_mS73PCKeCMqqeaWG_HdhtDH93kLKeXLIwjsZD2CUt64bzuiFvUhtDShGGk5kSve9Alw50q0sHhf16lO66Cfo38vjpBfh5ADYpmyc4ASZmZ0c4qsTrFOEpt2sTNXj2HwFel04</recordid><startdate>2002</startdate><enddate>2002</enddate><creator>Tanaka, Toshiya</creator><creator>Takeno, Tetsu</creator><creator>Watanabe, Yuichiro</creator><creator>Uchiyama, Yasutoshi</creator><creator>Murakami, Takeshi</creator><creator>Yamashita, Hisahiko</creator><creator>Suzuki, Akifumi</creator><creator>Aoi, Rie</creator><creator>Iwanari, Hiroko</creator><creator>Jiang, Shu-Ying</creator><creator>Naito, Makoto</creator><creator>Tachibana, Keisuke</creator><creator>Doi, Takefumi</creator><creator>Shulman, Andrew I.</creator><creator>Mangelsdorf, David J.</creator><creator>Reiter, Raphael</creator><creator>Auwerx, Johan</creator><creator>Hamakubo, Takao</creator><creator>Kodama, Tatsuhiko</creator><general>Japan Atherosclerosis Society</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope></search><sort><creationdate>2002</creationdate><title>The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs)</title><author>Tanaka, Toshiya ; Takeno, Tetsu ; Watanabe, Yuichiro ; Uchiyama, Yasutoshi ; Murakami, Takeshi ; Yamashita, Hisahiko ; Suzuki, Akifumi ; Aoi, Rie ; Iwanari, Hiroko ; Jiang, Shu-Ying ; Naito, Makoto ; Tachibana, Keisuke ; Doi, Takefumi ; Shulman, Andrew I. ; Mangelsdorf, David J. ; Reiter, Raphael ; Auwerx, Johan ; Hamakubo, Takao ; Kodama, Tatsuhiko</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5103-42c42bc57d47447f9a3b790598c7f6518397b79d500e8290c051fdb6f64b5c843</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - genetics</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Baculovirus</topic><topic>Blotting, Western</topic><topic>CHO Cells</topic><topic>Cricetinae</topic><topic>Electrophoretic Mobility Shift Assay</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Female</topic><topic>Gp64-fusion protein</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Life Sciences</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Monoclonal antibody (Mab)</topic><topic>Nucleopolyhedrovirus - genetics</topic><topic>Peroxisome proliferator-activated receptors (PPARs)</topic><topic>Receptors, Cytoplasmic and Nuclear - immunology</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - immunology</topic><topic>Spodoptera</topic><topic>Transcription Factors - immunology</topic><toplevel>online_resources</toplevel><creatorcontrib>Tanaka, Toshiya</creatorcontrib><creatorcontrib>Takeno, Tetsu</creatorcontrib><creatorcontrib>Watanabe, Yuichiro</creatorcontrib><creatorcontrib>Uchiyama, Yasutoshi</creatorcontrib><creatorcontrib>Murakami, Takeshi</creatorcontrib><creatorcontrib>Yamashita, Hisahiko</creatorcontrib><creatorcontrib>Suzuki, Akifumi</creatorcontrib><creatorcontrib>Aoi, Rie</creatorcontrib><creatorcontrib>Iwanari, Hiroko</creatorcontrib><creatorcontrib>Jiang, Shu-Ying</creatorcontrib><creatorcontrib>Naito, Makoto</creatorcontrib><creatorcontrib>Tachibana, Keisuke</creatorcontrib><creatorcontrib>Doi, Takefumi</creatorcontrib><creatorcontrib>Shulman, Andrew I.</creatorcontrib><creatorcontrib>Mangelsdorf, David J.</creatorcontrib><creatorcontrib>Reiter, Raphael</creatorcontrib><creatorcontrib>Auwerx, Johan</creatorcontrib><creatorcontrib>Hamakubo, Takao</creatorcontrib><creatorcontrib>Kodama, Tatsuhiko</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>Journal of Atherosclerosis and Thrombosis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tanaka, Toshiya</au><au>Takeno, Tetsu</au><au>Watanabe, Yuichiro</au><au>Uchiyama, Yasutoshi</au><au>Murakami, Takeshi</au><au>Yamashita, Hisahiko</au><au>Suzuki, Akifumi</au><au>Aoi, Rie</au><au>Iwanari, Hiroko</au><au>Jiang, Shu-Ying</au><au>Naito, Makoto</au><au>Tachibana, Keisuke</au><au>Doi, Takefumi</au><au>Shulman, Andrew I.</au><au>Mangelsdorf, David J.</au><au>Reiter, Raphael</au><au>Auwerx, Johan</au><au>Hamakubo, Takao</au><au>Kodama, Tatsuhiko</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs)</atitle><jtitle>Journal of Atherosclerosis and Thrombosis</jtitle><addtitle>JAT</addtitle><date>2002</date><risdate>2002</risdate><volume>9</volume><issue>5</issue><spage>233</spage><epage>242</epage><pages>233-242</pages><issn>1340-3478</issn><eissn>1880-3873</eissn><abstract>Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.</abstract><cop>Japan</cop><pub>Japan Atherosclerosis Society</pub><pmid>12409633</pmid><doi>10.5551/jat.9.233</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1340-3478 |
| ispartof | Journal of Atherosclerosis and Thrombosis, 2002, Vol.9(5), pp.233-242 |
| issn | 1340-3478 1880-3873 |
| language | eng |
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| source | MEDLINE; J-STAGE (Japan Science & Technology Information Aggregator, Electronic) Freely Available Titles - Japanese; EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Antibodies, Monoclonal - genetics Antibodies, Monoclonal - immunology Baculovirus Blotting, Western CHO Cells Cricetinae Electrophoretic Mobility Shift Assay Enzyme-Linked Immunosorbent Assay Female Gp64-fusion protein Humans Immunohistochemistry Life Sciences Mice Mice, Inbred BALB C Monoclonal antibody (Mab) Nucleopolyhedrovirus - genetics Peroxisome proliferator-activated receptors (PPARs) Receptors, Cytoplasmic and Nuclear - immunology Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - immunology Spodoptera Transcription Factors - immunology |
| title | The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs) |
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