Production and characterization of a colon cancer-specific immunotoxin based on the fungal ribotoxin α-sarcin
A single-chain fusion protein that directed the cytolytic activity of α-sarcin to A33 tumor antigen expressing cells was constructed and shown to effectively kill targeted cells. Glycoprotein A33 (GPA33) is a well-known colon cancer marker and a humanized antibody against it was used to target the α...
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| Veröffentlicht in: | Protein engineering, design and selection design and selection, 2012-08, Vol.25 (8), p.425-435 |
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| creator | Carreras-Sangrà, Nelson Tomé-Amat, Jaime García-Ortega, Lucía Batt, Carl A. Oñaderra, Mercedes Martínez-del-Pozo, Álvaro Gavilanes, José G. Lacadena, Javier |
| description | A single-chain fusion protein that directed the cytolytic activity of α-sarcin to A33 tumor antigen expressing cells was constructed and shown to effectively kill targeted cells. Glycoprotein A33 (GPA33) is a well-known colon cancer marker and a humanized antibody against it was used to target the α-sarcin. The fungal ribotoxin α-sarcin is one of the most potent and specific toxins known. It is small, protease resistant, thermostable and highly efficient towards the inactivation of ribosomes. This work describes the production and characterization of an immunotoxin resulting from fusing the single-chain variable fragment (scFv) of the monoclonal antibody that targets GPA33 to fungal α-sarcin. This chimeric protein (scFvA33αsarcin), produced in Pichia pastoris and purified in high yield was proven to be properly folded, active, specific and stable. It showed high specific toxicity against GPA33-positive tumoral cell lines providing scientific evidence to sustain that scFvA33αsarcin is a good immunotherapeutic candidate against GPA33-positive colon carcinomas. |
| doi_str_mv | 10.1093/protein/gzs032 |
| format | Article |
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Glycoprotein A33 (GPA33) is a well-known colon cancer marker and a humanized antibody against it was used to target the α-sarcin. The fungal ribotoxin α-sarcin is one of the most potent and specific toxins known. It is small, protease resistant, thermostable and highly efficient towards the inactivation of ribosomes. This work describes the production and characterization of an immunotoxin resulting from fusing the single-chain variable fragment (scFv) of the monoclonal antibody that targets GPA33 to fungal α-sarcin. This chimeric protein (scFvA33αsarcin), produced in Pichia pastoris and purified in high yield was proven to be properly folded, active, specific and stable. It showed high specific toxicity against GPA33-positive tumoral cell lines providing scientific evidence to sustain that scFvA33αsarcin is a good immunotherapeutic candidate against GPA33-positive colon carcinomas.</description><identifier>ISSN: 1741-0126</identifier><identifier>EISSN: 1741-0134</identifier><identifier>DOI: 10.1093/protein/gzs032</identifier><identifier>PMID: 22718791</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Cell Line, Tumor ; Colonic Neoplasms - drug therapy ; Colonic Neoplasms - immunology ; Colonic Neoplasms - metabolism ; Endoribonucleases - chemistry ; Endoribonucleases - genetics ; Endoribonucleases - metabolism ; Flow Cytometry ; Fungal Proteins - chemistry ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Humans ; Immunotoxins - chemistry ; Immunotoxins - genetics ; Immunotoxins - metabolism ; Membrane Glycoproteins - chemistry ; Membrane Glycoproteins - genetics ; Membrane Glycoproteins - metabolism ; Microscopy, Fluorescence ; Pichia - genetics ; Recombinant Fusion Proteins - chemistry ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - isolation & purification ; Recombinant Fusion Proteins - metabolism ; Single-Chain Antibodies - chemistry ; Single-Chain Antibodies - genetics ; Single-Chain Antibodies - metabolism</subject><ispartof>Protein engineering, design and selection, 2012-08, Vol.25 (8), p.425-435</ispartof><rights>The Author 2012. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com 2012</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1584,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22718791$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carreras-Sangrà, Nelson</creatorcontrib><creatorcontrib>Tomé-Amat, Jaime</creatorcontrib><creatorcontrib>García-Ortega, Lucía</creatorcontrib><creatorcontrib>Batt, Carl A.</creatorcontrib><creatorcontrib>Oñaderra, Mercedes</creatorcontrib><creatorcontrib>Martínez-del-Pozo, Álvaro</creatorcontrib><creatorcontrib>Gavilanes, José G.</creatorcontrib><creatorcontrib>Lacadena, Javier</creatorcontrib><title>Production and characterization of a colon cancer-specific immunotoxin based on the fungal ribotoxin α-sarcin</title><title>Protein engineering, design and selection</title><addtitle>Protein Eng Des Sel</addtitle><description>A single-chain fusion protein that directed the cytolytic activity of α-sarcin to A33 tumor antigen expressing cells was constructed and shown to effectively kill targeted cells. Glycoprotein A33 (GPA33) is a well-known colon cancer marker and a humanized antibody against it was used to target the α-sarcin. The fungal ribotoxin α-sarcin is one of the most potent and specific toxins known. It is small, protease resistant, thermostable and highly efficient towards the inactivation of ribosomes. This work describes the production and characterization of an immunotoxin resulting from fusing the single-chain variable fragment (scFv) of the monoclonal antibody that targets GPA33 to fungal α-sarcin. This chimeric protein (scFvA33αsarcin), produced in Pichia pastoris and purified in high yield was proven to be properly folded, active, specific and stable. It showed high specific toxicity against GPA33-positive tumoral cell lines providing scientific evidence to sustain that scFvA33αsarcin is a good immunotherapeutic candidate against GPA33-positive colon carcinomas.</description><subject>Cell Line, Tumor</subject><subject>Colonic Neoplasms - drug therapy</subject><subject>Colonic Neoplasms - immunology</subject><subject>Colonic Neoplasms - metabolism</subject><subject>Endoribonucleases - chemistry</subject><subject>Endoribonucleases - genetics</subject><subject>Endoribonucleases - metabolism</subject><subject>Flow Cytometry</subject><subject>Fungal Proteins - chemistry</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - metabolism</subject><subject>Humans</subject><subject>Immunotoxins - chemistry</subject><subject>Immunotoxins - genetics</subject><subject>Immunotoxins - metabolism</subject><subject>Membrane Glycoproteins - chemistry</subject><subject>Membrane Glycoproteins - genetics</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>Microscopy, Fluorescence</subject><subject>Pichia - genetics</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - isolation & purification</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Single-Chain Antibodies - chemistry</subject><subject>Single-Chain Antibodies - genetics</subject><subject>Single-Chain Antibodies - metabolism</subject><issn>1741-0126</issn><issn>1741-0134</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kM1OwzAQhC0EoqVw5Yh85RC6_omTHFHFn1QJDnCONrbTGjV25CQS9K14EZ6JQEtPO9oZjTQfIZcMbhgUYt7G0Fvn56ttB4IfkSnLJEuACXl80FxNyFnXvQNwlTF2SiacZyzPCjYl_iUGM-jeBU_RG6rXGFH3Nrot_j1DTZHqsBmlRq9tTLrWalc7TV3TDD704cN5WmFnDR1D_drSevAr3NDoqr37_ZV0GLXz5-Skxk1nL_Z3Rt7u714Xj8ny-eFpcbtMAlPQJ5IraSGTwLkCU9RoDVeVgNTkFRcCKiW0xYKD5FJaZbkReYp5zVMzDjSFmJGrXW87VI01ZRtdg_Gz_B8-Bq53gTC0B5dB-Qu13EMtd1DFD83ybIo</recordid><startdate>201208</startdate><enddate>201208</enddate><creator>Carreras-Sangrà, Nelson</creator><creator>Tomé-Amat, Jaime</creator><creator>García-Ortega, Lucía</creator><creator>Batt, Carl A.</creator><creator>Oñaderra, Mercedes</creator><creator>Martínez-del-Pozo, Álvaro</creator><creator>Gavilanes, José G.</creator><creator>Lacadena, Javier</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope></search><sort><creationdate>201208</creationdate><title>Production and characterization of a colon cancer-specific immunotoxin based on the fungal ribotoxin α-sarcin</title><author>Carreras-Sangrà, Nelson ; Tomé-Amat, Jaime ; García-Ortega, Lucía ; Batt, Carl A. ; Oñaderra, Mercedes ; Martínez-del-Pozo, Álvaro ; Gavilanes, José G. ; Lacadena, Javier</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-o160t-4264e07402260d9faed26b305d8b2330b63cea9204244e6e2d385a8f25d711d93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Cell Line, Tumor</topic><topic>Colonic Neoplasms - drug therapy</topic><topic>Colonic Neoplasms - immunology</topic><topic>Colonic Neoplasms - metabolism</topic><topic>Endoribonucleases - chemistry</topic><topic>Endoribonucleases - genetics</topic><topic>Endoribonucleases - metabolism</topic><topic>Flow Cytometry</topic><topic>Fungal Proteins - chemistry</topic><topic>Fungal Proteins - genetics</topic><topic>Fungal Proteins - metabolism</topic><topic>Humans</topic><topic>Immunotoxins - chemistry</topic><topic>Immunotoxins - genetics</topic><topic>Immunotoxins - metabolism</topic><topic>Membrane Glycoproteins - chemistry</topic><topic>Membrane Glycoproteins - genetics</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>Microscopy, Fluorescence</topic><topic>Pichia - genetics</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - isolation & purification</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Single-Chain Antibodies - chemistry</topic><topic>Single-Chain Antibodies - genetics</topic><topic>Single-Chain Antibodies - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Carreras-Sangrà, Nelson</creatorcontrib><creatorcontrib>Tomé-Amat, Jaime</creatorcontrib><creatorcontrib>García-Ortega, Lucía</creatorcontrib><creatorcontrib>Batt, Carl A.</creatorcontrib><creatorcontrib>Oñaderra, Mercedes</creatorcontrib><creatorcontrib>Martínez-del-Pozo, Álvaro</creatorcontrib><creatorcontrib>Gavilanes, José G.</creatorcontrib><creatorcontrib>Lacadena, Javier</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><jtitle>Protein engineering, design and selection</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Carreras-Sangrà, Nelson</au><au>Tomé-Amat, Jaime</au><au>García-Ortega, Lucía</au><au>Batt, Carl A.</au><au>Oñaderra, Mercedes</au><au>Martínez-del-Pozo, Álvaro</au><au>Gavilanes, José G.</au><au>Lacadena, Javier</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production and characterization of a colon cancer-specific immunotoxin based on the fungal ribotoxin α-sarcin</atitle><jtitle>Protein engineering, design and selection</jtitle><addtitle>Protein Eng Des Sel</addtitle><date>2012-08</date><risdate>2012</risdate><volume>25</volume><issue>8</issue><spage>425</spage><epage>435</epage><pages>425-435</pages><issn>1741-0126</issn><eissn>1741-0134</eissn><abstract>A single-chain fusion protein that directed the cytolytic activity of α-sarcin to A33 tumor antigen expressing cells was constructed and shown to effectively kill targeted cells. Glycoprotein A33 (GPA33) is a well-known colon cancer marker and a humanized antibody against it was used to target the α-sarcin. The fungal ribotoxin α-sarcin is one of the most potent and specific toxins known. It is small, protease resistant, thermostable and highly efficient towards the inactivation of ribosomes. This work describes the production and characterization of an immunotoxin resulting from fusing the single-chain variable fragment (scFv) of the monoclonal antibody that targets GPA33 to fungal α-sarcin. This chimeric protein (scFvA33αsarcin), produced in Pichia pastoris and purified in high yield was proven to be properly folded, active, specific and stable. It showed high specific toxicity against GPA33-positive tumoral cell lines providing scientific evidence to sustain that scFvA33αsarcin is a good immunotherapeutic candidate against GPA33-positive colon carcinomas.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>22718791</pmid><doi>10.1093/protein/gzs032</doi><tpages>11</tpages></addata></record> |
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| source | MEDLINE; Oxford Journals (Firm); Alma/SFX Local Collection; EZB Electronic Journals Library |
| subjects | Cell Line, Tumor Colonic Neoplasms - drug therapy Colonic Neoplasms - immunology Colonic Neoplasms - metabolism Endoribonucleases - chemistry Endoribonucleases - genetics Endoribonucleases - metabolism Flow Cytometry Fungal Proteins - chemistry Fungal Proteins - genetics Fungal Proteins - metabolism Humans Immunotoxins - chemistry Immunotoxins - genetics Immunotoxins - metabolism Membrane Glycoproteins - chemistry Membrane Glycoproteins - genetics Membrane Glycoproteins - metabolism Microscopy, Fluorescence Pichia - genetics Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - isolation & purification Recombinant Fusion Proteins - metabolism Single-Chain Antibodies - chemistry Single-Chain Antibodies - genetics Single-Chain Antibodies - metabolism |
| title | Production and characterization of a colon cancer-specific immunotoxin based on the fungal ribotoxin α-sarcin |
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