In situ phage screening. A method for identification of subnanogram tissue components in situ
We have established a novel method, in situ phage screening (ISPS), to identify proteins in tissue microstructures. The method is based on the selection of repertoires of phage-displayed antibody fragments with small samples of tissues microdissected using a laser. Using a human muscle frozen sectio...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 2002-08, Vol.277 (33), p.30382-30387 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 30387 |
|---|---|
| container_issue | 33 |
| container_start_page | 30382 |
| container_title | The Journal of biological chemistry |
| container_volume | 277 |
| creator | Tanaka, Torahiko Ito, Takashi Furuta, Masaru Eguchi, Chikashi Toda, Hiroyuki Wakabayashi-Takai, Eriko Kaneko, Kiyotoshi |
| description | We have established a novel method, in situ phage screening (ISPS), to identify proteins in tissue microstructures. The method is based on the selection of repertoires of phage-displayed antibody fragments with small samples of tissues microdissected using a laser. Using a human muscle frozen section with an area of 4800 microm2 as a model target, we successfully selected monoclonal antibody fragments directed against three major (myosin heavy chain, actin, and tropomyosin-alpha) and one minor (alpha-actinin 2) muscle constituent proteins. These proteins were present in the sample in amounts less than one nanogram, and the antibodies were used to visualize the proteins in situ. This shows that the use of ISPS can obtain monoclonal antibodies for histochemical and biochemical purposes against minute amounts of proteins from microstructures with no requirement for large amounts of samples or biochemical efforts. |
| doi_str_mv | 10.1074/jbc.M203547200 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_71991394</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>71991394</sourcerecordid><originalsourceid>FETCH-LOGICAL-p238t-d5910df2bf2cd1c46fb613cada1d3f20586c9926b642f3780549f31dce216b483</originalsourceid><addsrcrecordid>eNqF0D1PwzAYBGAPIFoKKyPyxJbi13YSe6wqKJWKWGBEkeOP1lVjh9gZ-PcUtczccsujGw6hOyBzIDV_3Ld6_koJK3lNCblAU0IoFJKWYoKuU9qTY7iEKzQBSrigBKbocx1w8nnE_U5tLU56sDb4sJ3jBe5s3kWDXRywNzZk77xW2ceAo8NpbIMKcTuoDmef0mixjl0fwxEm7E-rN-jSqUOyt-eeoY_np_flS7F5W62Xi03RUyZyYUoJxDjaOqoNaF65tgKmlVFgmKOkFJWWklZtxaljtSAll46B0ZZC1XLBZujhtNsP8Wu0KTedT9oeDirYOKamBimBSf4vBFGKkrNfeH-GY9tZ0_SD79Tw3fw9x34ATuFuqw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18585434</pqid></control><display><type>article</type><title>In situ phage screening. A method for identification of subnanogram tissue components in situ</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Tanaka, Torahiko ; Ito, Takashi ; Furuta, Masaru ; Eguchi, Chikashi ; Toda, Hiroyuki ; Wakabayashi-Takai, Eriko ; Kaneko, Kiyotoshi</creator><creatorcontrib>Tanaka, Torahiko ; Ito, Takashi ; Furuta, Masaru ; Eguchi, Chikashi ; Toda, Hiroyuki ; Wakabayashi-Takai, Eriko ; Kaneko, Kiyotoshi</creatorcontrib><description>We have established a novel method, in situ phage screening (ISPS), to identify proteins in tissue microstructures. The method is based on the selection of repertoires of phage-displayed antibody fragments with small samples of tissues microdissected using a laser. Using a human muscle frozen section with an area of 4800 microm2 as a model target, we successfully selected monoclonal antibody fragments directed against three major (myosin heavy chain, actin, and tropomyosin-alpha) and one minor (alpha-actinin 2) muscle constituent proteins. These proteins were present in the sample in amounts less than one nanogram, and the antibodies were used to visualize the proteins in situ. This shows that the use of ISPS can obtain monoclonal antibodies for histochemical and biochemical purposes against minute amounts of proteins from microstructures with no requirement for large amounts of samples or biochemical efforts.</description><identifier>ISSN: 0021-9258</identifier><identifier>DOI: 10.1074/jbc.M203547200</identifier><identifier>PMID: 12048201</identifier><language>eng</language><publisher>United States</publisher><subject>Antigens - genetics ; Bacteriophages - genetics ; Base Sequence ; DNA Primers ; Humans ; Immunoglobulin Fragments - genetics ; Muscle, Skeletal - metabolism</subject><ispartof>The Journal of biological chemistry, 2002-08, Vol.277 (33), p.30382-30387</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12048201$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tanaka, Torahiko</creatorcontrib><creatorcontrib>Ito, Takashi</creatorcontrib><creatorcontrib>Furuta, Masaru</creatorcontrib><creatorcontrib>Eguchi, Chikashi</creatorcontrib><creatorcontrib>Toda, Hiroyuki</creatorcontrib><creatorcontrib>Wakabayashi-Takai, Eriko</creatorcontrib><creatorcontrib>Kaneko, Kiyotoshi</creatorcontrib><title>In situ phage screening. A method for identification of subnanogram tissue components in situ</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>We have established a novel method, in situ phage screening (ISPS), to identify proteins in tissue microstructures. The method is based on the selection of repertoires of phage-displayed antibody fragments with small samples of tissues microdissected using a laser. Using a human muscle frozen section with an area of 4800 microm2 as a model target, we successfully selected monoclonal antibody fragments directed against three major (myosin heavy chain, actin, and tropomyosin-alpha) and one minor (alpha-actinin 2) muscle constituent proteins. These proteins were present in the sample in amounts less than one nanogram, and the antibodies were used to visualize the proteins in situ. This shows that the use of ISPS can obtain monoclonal antibodies for histochemical and biochemical purposes against minute amounts of proteins from microstructures with no requirement for large amounts of samples or biochemical efforts.</description><subject>Antigens - genetics</subject><subject>Bacteriophages - genetics</subject><subject>Base Sequence</subject><subject>DNA Primers</subject><subject>Humans</subject><subject>Immunoglobulin Fragments - genetics</subject><subject>Muscle, Skeletal - metabolism</subject><issn>0021-9258</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0D1PwzAYBGAPIFoKKyPyxJbi13YSe6wqKJWKWGBEkeOP1lVjh9gZ-PcUtczccsujGw6hOyBzIDV_3Ld6_koJK3lNCblAU0IoFJKWYoKuU9qTY7iEKzQBSrigBKbocx1w8nnE_U5tLU56sDb4sJ3jBe5s3kWDXRywNzZk77xW2ceAo8NpbIMKcTuoDmef0mixjl0fwxEm7E-rN-jSqUOyt-eeoY_np_flS7F5W62Xi03RUyZyYUoJxDjaOqoNaF65tgKmlVFgmKOkFJWWklZtxaljtSAll46B0ZZC1XLBZujhtNsP8Wu0KTedT9oeDirYOKamBimBSf4vBFGKkrNfeH-GY9tZ0_SD79Tw3fw9x34ATuFuqw</recordid><startdate>20020816</startdate><enddate>20020816</enddate><creator>Tanaka, Torahiko</creator><creator>Ito, Takashi</creator><creator>Furuta, Masaru</creator><creator>Eguchi, Chikashi</creator><creator>Toda, Hiroyuki</creator><creator>Wakabayashi-Takai, Eriko</creator><creator>Kaneko, Kiyotoshi</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20020816</creationdate><title>In situ phage screening. A method for identification of subnanogram tissue components in situ</title><author>Tanaka, Torahiko ; Ito, Takashi ; Furuta, Masaru ; Eguchi, Chikashi ; Toda, Hiroyuki ; Wakabayashi-Takai, Eriko ; Kaneko, Kiyotoshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p238t-d5910df2bf2cd1c46fb613cada1d3f20586c9926b642f3780549f31dce216b483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Antigens - genetics</topic><topic>Bacteriophages - genetics</topic><topic>Base Sequence</topic><topic>DNA Primers</topic><topic>Humans</topic><topic>Immunoglobulin Fragments - genetics</topic><topic>Muscle, Skeletal - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tanaka, Torahiko</creatorcontrib><creatorcontrib>Ito, Takashi</creatorcontrib><creatorcontrib>Furuta, Masaru</creatorcontrib><creatorcontrib>Eguchi, Chikashi</creatorcontrib><creatorcontrib>Toda, Hiroyuki</creatorcontrib><creatorcontrib>Wakabayashi-Takai, Eriko</creatorcontrib><creatorcontrib>Kaneko, Kiyotoshi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tanaka, Torahiko</au><au>Ito, Takashi</au><au>Furuta, Masaru</au><au>Eguchi, Chikashi</au><au>Toda, Hiroyuki</au><au>Wakabayashi-Takai, Eriko</au><au>Kaneko, Kiyotoshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In situ phage screening. A method for identification of subnanogram tissue components in situ</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2002-08-16</date><risdate>2002</risdate><volume>277</volume><issue>33</issue><spage>30382</spage><epage>30387</epage><pages>30382-30387</pages><issn>0021-9258</issn><abstract>We have established a novel method, in situ phage screening (ISPS), to identify proteins in tissue microstructures. The method is based on the selection of repertoires of phage-displayed antibody fragments with small samples of tissues microdissected using a laser. Using a human muscle frozen section with an area of 4800 microm2 as a model target, we successfully selected monoclonal antibody fragments directed against three major (myosin heavy chain, actin, and tropomyosin-alpha) and one minor (alpha-actinin 2) muscle constituent proteins. These proteins were present in the sample in amounts less than one nanogram, and the antibodies were used to visualize the proteins in situ. This shows that the use of ISPS can obtain monoclonal antibodies for histochemical and biochemical purposes against minute amounts of proteins from microstructures with no requirement for large amounts of samples or biochemical efforts.</abstract><cop>United States</cop><pmid>12048201</pmid><doi>10.1074/jbc.M203547200</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 2002-08, Vol.277 (33), p.30382-30387 |
| issn | 0021-9258 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_71991394 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Antigens - genetics Bacteriophages - genetics Base Sequence DNA Primers Humans Immunoglobulin Fragments - genetics Muscle, Skeletal - metabolism |
| title | In situ phage screening. A method for identification of subnanogram tissue components in situ |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-24T07%3A02%3A39IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=In%20situ%20phage%20screening.%20A%20method%20for%20identification%20of%20subnanogram%20tissue%20components%20in%20situ&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Tanaka,%20Torahiko&rft.date=2002-08-16&rft.volume=277&rft.issue=33&rft.spage=30382&rft.epage=30387&rft.pages=30382-30387&rft.issn=0021-9258&rft_id=info:doi/10.1074/jbc.M203547200&rft_dat=%3Cproquest_pubme%3E71991394%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=18585434&rft_id=info:pmid/12048201&rfr_iscdi=true |