Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies Using a Phage Display Library
A new single-chain variable fragment (scFv) antibody library was generated and human serum albumin (HSA)-specific clones were characterized to investigate the usefulness of porcine antibodies. Phage libraries were developed from pigs immunized with the model antigen HSA. The library size was 1.5×10...
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| Veröffentlicht in: | Monoclonal antibodies in immunodiagnosis and immunotherapy 2014-02, Vol.33 (1), p.42-48 |
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| container_title | Monoclonal antibodies in immunodiagnosis and immunotherapy |
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| creator | Muraoka, Junko Ozawa, Takuya Enomoto, Yurie Kiyose, Norihiko Imamura, Ayana Arima, Kazunari Nakayama, Hiroshi Ito, Yuji |
| description | A new single-chain variable fragment (scFv) antibody library was generated and human serum albumin (HSA)-specific clones were characterized to investigate the usefulness of porcine antibodies. Phage libraries were developed from pigs immunized with the model antigen HSA. The library size was 1.5×10
7
for kappa (V
L
) and 1.4×10
7
for lambda fragments. Eight HSA-specific clones from the kappa library and one clone from the lambda library were isolated using affinity selection. The binding specificity of these clones was confirmed using a phage enzyme-linked immunosorbent assay (ELISA). The scFvs were expressed in
Escherichia coli
and purified from the periplasm fraction for further investigation. Based on the results of ELISA and Western blot analysis, four scFv clones with high activity and high yield were selected and purified. The purified scFvs from four of the nine clones exhibited an approximate K
D
of 10
−8
M. This is the first report describing isolation of HSA-specific porcine scFv antibodies from an antibody phage library and characterization of their binding properties. |
| doi_str_mv | 10.1089/mab.2013.0068 |
| format | Article |
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7
for kappa (V
L
) and 1.4×10
7
for lambda fragments. Eight HSA-specific clones from the kappa library and one clone from the lambda library were isolated using affinity selection. The binding specificity of these clones was confirmed using a phage enzyme-linked immunosorbent assay (ELISA). The scFvs were expressed in
Escherichia coli
and purified from the periplasm fraction for further investigation. Based on the results of ELISA and Western blot analysis, four scFv clones with high activity and high yield were selected and purified. The purified scFvs from four of the nine clones exhibited an approximate K
D
of 10
−8
M. This is the first report describing isolation of HSA-specific porcine scFv antibodies from an antibody phage library and characterization of their binding properties.</description><identifier>ISSN: 2167-9436</identifier><identifier>EISSN: 2167-9436</identifier><identifier>DOI: 10.1089/mab.2013.0068</identifier><identifier>PMID: 24555936</identifier><language>eng</language><publisher>United States: Mary Ann Liebert, Inc</publisher><subject>Amino Acid Sequence ; Animals ; Antibody Specificity - immunology ; Cell Surface Display Techniques ; Enzyme-Linked Immunosorbent Assay ; Gene Expression - immunology ; Humans ; Molecular Sequence Data ; Original Articles ; Peptide Library ; Serum Albumin - immunology ; Single-Chain Antibodies - immunology ; Sus scrofa - immunology</subject><ispartof>Monoclonal antibodies in immunodiagnosis and immunotherapy, 2014-02, Vol.33 (1), p.42-48</ispartof><rights>2014, Mary Ann Liebert, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c318t-107edead0928654c42230a928df785c0142cce2e1ab7dbe08dae3eeaf7dcd3f23</citedby><cites>FETCH-LOGICAL-c318t-107edead0928654c42230a928df785c0142cce2e1ab7dbe08dae3eeaf7dcd3f23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24555936$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Muraoka, Junko</creatorcontrib><creatorcontrib>Ozawa, Takuya</creatorcontrib><creatorcontrib>Enomoto, Yurie</creatorcontrib><creatorcontrib>Kiyose, Norihiko</creatorcontrib><creatorcontrib>Imamura, Ayana</creatorcontrib><creatorcontrib>Arima, Kazunari</creatorcontrib><creatorcontrib>Nakayama, Hiroshi</creatorcontrib><creatorcontrib>Ito, Yuji</creatorcontrib><title>Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies Using a Phage Display Library</title><title>Monoclonal antibodies in immunodiagnosis and immunotherapy</title><addtitle>Monoclon Antib Immunodiagn Immunother</addtitle><description>A new single-chain variable fragment (scFv) antibody library was generated and human serum albumin (HSA)-specific clones were characterized to investigate the usefulness of porcine antibodies. Phage libraries were developed from pigs immunized with the model antigen HSA. The library size was 1.5×10
7
for kappa (V
L
) and 1.4×10
7
for lambda fragments. Eight HSA-specific clones from the kappa library and one clone from the lambda library were isolated using affinity selection. The binding specificity of these clones was confirmed using a phage enzyme-linked immunosorbent assay (ELISA). The scFvs were expressed in
Escherichia coli
and purified from the periplasm fraction for further investigation. Based on the results of ELISA and Western blot analysis, four scFv clones with high activity and high yield were selected and purified. The purified scFvs from four of the nine clones exhibited an approximate K
D
of 10
−8
M. This is the first report describing isolation of HSA-specific porcine scFv antibodies from an antibody phage library and characterization of their binding properties.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibody Specificity - immunology</subject><subject>Cell Surface Display Techniques</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Gene Expression - immunology</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><subject>Original Articles</subject><subject>Peptide Library</subject><subject>Serum Albumin - immunology</subject><subject>Single-Chain Antibodies - immunology</subject><subject>Sus scrofa - immunology</subject><issn>2167-9436</issn><issn>2167-9436</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtLxDAQh4MoKurRq-TopWsefR6X1VVhwYXVc5kmU420aU1aYf3rTV0Vb85lHnz8GD5CzjmbcZYXVy1UM8G4nDGW5nvkWPA0i4pYpvt_5iNy5v0rC5VxwWVySI5EnCRJIdNjMmywQTWYzlKwmi5ewIEa0JkP-Dp2Nb0bW7B0g25s6bypxtbYyPeoTG0UXXdOGYvUq-U7ndvBVJ026OmTN_aZAl2_wDPSa-P7BrZ0ZSoHbntKDmpoPJ599xPytLx5XNxFq4fb-8V8FSnJ8yHiLEONoFkh8jSJVSyEZBAWXWd5ohiPhVIokEOV6QpZrgElItSZVlrWQp6Qy11u77q3Ef1QtsYrbBqw2I2-5Elwl_FQAY12qHKd9w7rsnemDb-WnJWT6zK4LifX5eQ68Bff0WPVov6lf8wGQO6A6QzWNgYrdMM_sZ-4coyY</recordid><startdate>20140201</startdate><enddate>20140201</enddate><creator>Muraoka, Junko</creator><creator>Ozawa, Takuya</creator><creator>Enomoto, Yurie</creator><creator>Kiyose, Norihiko</creator><creator>Imamura, Ayana</creator><creator>Arima, Kazunari</creator><creator>Nakayama, Hiroshi</creator><creator>Ito, Yuji</creator><general>Mary Ann Liebert, Inc</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></search><sort><creationdate>20140201</creationdate><title>Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies Using a Phage Display Library</title><author>Muraoka, Junko ; Ozawa, Takuya ; Enomoto, Yurie ; Kiyose, Norihiko ; Imamura, Ayana ; Arima, Kazunari ; Nakayama, Hiroshi ; Ito, Yuji</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c318t-107edead0928654c42230a928df785c0142cce2e1ab7dbe08dae3eeaf7dcd3f23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibody Specificity - immunology</topic><topic>Cell Surface Display Techniques</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Gene Expression - immunology</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><topic>Original Articles</topic><topic>Peptide Library</topic><topic>Serum Albumin - immunology</topic><topic>Single-Chain Antibodies - immunology</topic><topic>Sus scrofa - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Muraoka, Junko</creatorcontrib><creatorcontrib>Ozawa, Takuya</creatorcontrib><creatorcontrib>Enomoto, Yurie</creatorcontrib><creatorcontrib>Kiyose, Norihiko</creatorcontrib><creatorcontrib>Imamura, Ayana</creatorcontrib><creatorcontrib>Arima, Kazunari</creatorcontrib><creatorcontrib>Nakayama, Hiroshi</creatorcontrib><creatorcontrib>Ito, Yuji</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><jtitle>Monoclonal antibodies in immunodiagnosis and immunotherapy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Muraoka, Junko</au><au>Ozawa, Takuya</au><au>Enomoto, Yurie</au><au>Kiyose, Norihiko</au><au>Imamura, Ayana</au><au>Arima, Kazunari</au><au>Nakayama, Hiroshi</au><au>Ito, Yuji</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies Using a Phage Display Library</atitle><jtitle>Monoclonal antibodies in immunodiagnosis and immunotherapy</jtitle><addtitle>Monoclon Antib Immunodiagn Immunother</addtitle><date>2014-02-01</date><risdate>2014</risdate><volume>33</volume><issue>1</issue><spage>42</spage><epage>48</epage><pages>42-48</pages><issn>2167-9436</issn><eissn>2167-9436</eissn><abstract>A new single-chain variable fragment (scFv) antibody library was generated and human serum albumin (HSA)-specific clones were characterized to investigate the usefulness of porcine antibodies. Phage libraries were developed from pigs immunized with the model antigen HSA. The library size was 1.5×10
7
for kappa (V
L
) and 1.4×10
7
for lambda fragments. Eight HSA-specific clones from the kappa library and one clone from the lambda library were isolated using affinity selection. The binding specificity of these clones was confirmed using a phage enzyme-linked immunosorbent assay (ELISA). The scFvs were expressed in
Escherichia coli
and purified from the periplasm fraction for further investigation. Based on the results of ELISA and Western blot analysis, four scFv clones with high activity and high yield were selected and purified. The purified scFvs from four of the nine clones exhibited an approximate K
D
of 10
−8
M. This is the first report describing isolation of HSA-specific porcine scFv antibodies from an antibody phage library and characterization of their binding properties.</abstract><cop>United States</cop><pub>Mary Ann Liebert, Inc</pub><pmid>24555936</pmid><doi>10.1089/mab.2013.0068</doi><tpages>7</tpages></addata></record> |
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| ispartof | Monoclonal antibodies in immunodiagnosis and immunotherapy, 2014-02, Vol.33 (1), p.42-48 |
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| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence Animals Antibody Specificity - immunology Cell Surface Display Techniques Enzyme-Linked Immunosorbent Assay Gene Expression - immunology Humans Molecular Sequence Data Original Articles Peptide Library Serum Albumin - immunology Single-Chain Antibodies - immunology Sus scrofa - immunology |
| title | Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies Using a Phage Display Library |
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