Production and purification of melanoma gp100 antigen and polyclonal antibodies
gp100 is a melanoma-associated antigen found to carry immunogenic epitopes that can induce a CTL response against tumor cells. Production and purification of large quantities of this polypeptide may be important in the context of diagnosis and vaccinating against melanoma. To overcome the hydrophobi...
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| Veröffentlicht in: | Protein expression and purification 2004-04, Vol.34 (2), p.183-189 |
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| container_title | Protein expression and purification |
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| creator | Gelbart, Y Frankenburg, S Pinchasov, Y Krispel, S Eliahu, D Drize, O Morag, E Bartfeld, D Lotem, M Peretz, T Pitcovski, J |
| description | gp100 is a melanoma-associated antigen found to carry immunogenic epitopes that can induce a CTL response against tumor cells. Production and purification of large quantities of this polypeptide may be important in the context of diagnosis and vaccinating against melanoma. To overcome the hydrophobic nature of gp100, we cloned and expressed only a part of the protein, and obtained a hydrophilic recombinant polypeptide (HR-gp100) that contained most of the immunogenic peptides. High yield was achieved in an
Escherichia coli expression system. The protein was purified by AKTA Prime using anionic-columns. Polyclonal antibodies developed in chicken against HR-gp100 were efficient at detecting gp100 in melanoma cells, as determined by Western blot analysis and by immunohistochemistry. HR-gp100 can be used to develop a vaccine against melanoma. Antibodies to HR-gp100 may be used to detect tumors of melanocytic origin or to determine the level of gp100 expression in tumors prior to immunotherapy with the protein or one of its peptides. |
| doi_str_mv | 10.1016/j.pep.2003.12.006 |
| format | Article |
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Escherichia coli expression system. The protein was purified by AKTA Prime using anionic-columns. Polyclonal antibodies developed in chicken against HR-gp100 were efficient at detecting gp100 in melanoma cells, as determined by Western blot analysis and by immunohistochemistry. HR-gp100 can be used to develop a vaccine against melanoma. Antibodies to HR-gp100 may be used to detect tumors of melanocytic origin or to determine the level of gp100 expression in tumors prior to immunotherapy with the protein or one of its peptides.</description><identifier>ISSN: 1046-5928</identifier><identifier>EISSN: 1096-0279</identifier><identifier>DOI: 10.1016/j.pep.2003.12.006</identifier><identifier>PMID: 15003249</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Antibodies ; Antibodies - immunology ; Chickens - immunology ; Epitopes - immunology ; Escherichia coli - genetics ; gp100 ; gp100 Melanoma Antigen ; Humans ; Hydrophobic and Hydrophilic Interactions ; Immunohistochemistry ; Melanoma ; Melanoma - immunology ; Membrane Glycoproteins - isolation & purification ; Membrane Glycoproteins - metabolism ; Molecular Sequence Data ; Neoplasm Proteins - isolation & purification ; Neoplasm Proteins - metabolism ; Recombinant Proteins - immunology ; Tumor Cells, Cultured</subject><ispartof>Protein expression and purification, 2004-04, Vol.34 (2), p.183-189</ispartof><rights>2004 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-592b9ba08a5e08d5b8fc0c4716598f17be5edeab9f3f12598476522e168dcf523</citedby><cites>FETCH-LOGICAL-c417t-592b9ba08a5e08d5b8fc0c4716598f17be5edeab9f3f12598476522e168dcf523</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1046592804000087$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15003249$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gelbart, Y</creatorcontrib><creatorcontrib>Frankenburg, S</creatorcontrib><creatorcontrib>Pinchasov, Y</creatorcontrib><creatorcontrib>Krispel, S</creatorcontrib><creatorcontrib>Eliahu, D</creatorcontrib><creatorcontrib>Drize, O</creatorcontrib><creatorcontrib>Morag, E</creatorcontrib><creatorcontrib>Bartfeld, D</creatorcontrib><creatorcontrib>Lotem, M</creatorcontrib><creatorcontrib>Peretz, T</creatorcontrib><creatorcontrib>Pitcovski, J</creatorcontrib><title>Production and purification of melanoma gp100 antigen and polyclonal antibodies</title><title>Protein expression and purification</title><addtitle>Protein Expr Purif</addtitle><description>gp100 is a melanoma-associated antigen found to carry immunogenic epitopes that can induce a CTL response against tumor cells. Production and purification of large quantities of this polypeptide may be important in the context of diagnosis and vaccinating against melanoma. To overcome the hydrophobic nature of gp100, we cloned and expressed only a part of the protein, and obtained a hydrophilic recombinant polypeptide (HR-gp100) that contained most of the immunogenic peptides. High yield was achieved in an
Escherichia coli expression system. The protein was purified by AKTA Prime using anionic-columns. Polyclonal antibodies developed in chicken against HR-gp100 were efficient at detecting gp100 in melanoma cells, as determined by Western blot analysis and by immunohistochemistry. HR-gp100 can be used to develop a vaccine against melanoma. Antibodies to HR-gp100 may be used to detect tumors of melanocytic origin or to determine the level of gp100 expression in tumors prior to immunotherapy with the protein or one of its peptides.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies - immunology</subject><subject>Chickens - immunology</subject><subject>Epitopes - immunology</subject><subject>Escherichia coli - genetics</subject><subject>gp100</subject><subject>gp100 Melanoma Antigen</subject><subject>Humans</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>Immunohistochemistry</subject><subject>Melanoma</subject><subject>Melanoma - immunology</subject><subject>Membrane Glycoproteins - isolation & purification</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>Molecular Sequence Data</subject><subject>Neoplasm Proteins - isolation & purification</subject><subject>Neoplasm Proteins - metabolism</subject><subject>Recombinant Proteins - immunology</subject><subject>Tumor Cells, Cultured</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kElLBDEQhYMoLqM_wIvMydu0VZlOL3gScQNBD3oO6aQyZOjutEm34L83s4A3T1U8vveoeoxdImQIWNyss4GGjAMsM-QZQHHAThHqYgG8rA83e14sRM2rE3YW4xoAsQBxzE5QJA_P61P29h68mfTofD9XvZkPU3DWabUVvJ131Kred2q-GhAgIaNb0R717Y9ufa_ardx44yiesyOr2kgX-zljn48PH_fPi9e3p5f7u9eFzrEcN0c1daOgUoKgMqKprAadl1iIurJYNiTIkGpqu7TIk5aXheCcsKiMtoIvZ-x6lzsE_zVRHGXnoqY2XUt-irLEEnkFVQJxB-rgYwxk5RBcp8KPRJCbFuVaphblpkWJXKYWk-dqHz41HZk_x762BNzuAEovfjsKMmpHvSbjAulRGu_-if8FIY2CkQ</recordid><startdate>20040401</startdate><enddate>20040401</enddate><creator>Gelbart, Y</creator><creator>Frankenburg, S</creator><creator>Pinchasov, Y</creator><creator>Krispel, S</creator><creator>Eliahu, D</creator><creator>Drize, O</creator><creator>Morag, E</creator><creator>Bartfeld, D</creator><creator>Lotem, M</creator><creator>Peretz, T</creator><creator>Pitcovski, J</creator><general>Elsevier 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>20040401</creationdate><title>Production and purification of melanoma gp100 antigen and polyclonal antibodies</title><author>Gelbart, Y ; Frankenburg, S ; Pinchasov, Y ; Krispel, S ; Eliahu, D ; Drize, O ; Morag, E ; Bartfeld, D ; Lotem, M ; Peretz, T ; Pitcovski, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-592b9ba08a5e08d5b8fc0c4716598f17be5edeab9f3f12598476522e168dcf523</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies - immunology</topic><topic>Chickens - immunology</topic><topic>Epitopes - immunology</topic><topic>Escherichia coli - genetics</topic><topic>gp100</topic><topic>gp100 Melanoma Antigen</topic><topic>Humans</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>Immunohistochemistry</topic><topic>Melanoma</topic><topic>Melanoma - immunology</topic><topic>Membrane Glycoproteins - isolation & purification</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>Molecular Sequence Data</topic><topic>Neoplasm Proteins - isolation & purification</topic><topic>Neoplasm Proteins - metabolism</topic><topic>Recombinant Proteins - immunology</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gelbart, Y</creatorcontrib><creatorcontrib>Frankenburg, S</creatorcontrib><creatorcontrib>Pinchasov, Y</creatorcontrib><creatorcontrib>Krispel, S</creatorcontrib><creatorcontrib>Eliahu, D</creatorcontrib><creatorcontrib>Drize, O</creatorcontrib><creatorcontrib>Morag, E</creatorcontrib><creatorcontrib>Bartfeld, D</creatorcontrib><creatorcontrib>Lotem, M</creatorcontrib><creatorcontrib>Peretz, T</creatorcontrib><creatorcontrib>Pitcovski, J</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>Protein expression and purification</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gelbart, Y</au><au>Frankenburg, S</au><au>Pinchasov, Y</au><au>Krispel, S</au><au>Eliahu, D</au><au>Drize, O</au><au>Morag, E</au><au>Bartfeld, D</au><au>Lotem, M</au><au>Peretz, T</au><au>Pitcovski, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production and purification of melanoma gp100 antigen and polyclonal antibodies</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>2004-04-01</date><risdate>2004</risdate><volume>34</volume><issue>2</issue><spage>183</spage><epage>189</epage><pages>183-189</pages><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>gp100 is a melanoma-associated antigen found to carry immunogenic epitopes that can induce a CTL response against tumor cells. Production and purification of large quantities of this polypeptide may be important in the context of diagnosis and vaccinating against melanoma. To overcome the hydrophobic nature of gp100, we cloned and expressed only a part of the protein, and obtained a hydrophilic recombinant polypeptide (HR-gp100) that contained most of the immunogenic peptides. High yield was achieved in an
Escherichia coli expression system. The protein was purified by AKTA Prime using anionic-columns. Polyclonal antibodies developed in chicken against HR-gp100 were efficient at detecting gp100 in melanoma cells, as determined by Western blot analysis and by immunohistochemistry. HR-gp100 can be used to develop a vaccine against melanoma. Antibodies to HR-gp100 may be used to detect tumors of melanocytic origin or to determine the level of gp100 expression in tumors prior to immunotherapy with the protein or one of its peptides.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>15003249</pmid><doi>10.1016/j.pep.2003.12.006</doi><tpages>7</tpages></addata></record> |
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| subjects | Amino Acid Sequence Animals Antibodies Antibodies - immunology Chickens - immunology Epitopes - immunology Escherichia coli - genetics gp100 gp100 Melanoma Antigen Humans Hydrophobic and Hydrophilic Interactions Immunohistochemistry Melanoma Melanoma - immunology Membrane Glycoproteins - isolation & purification Membrane Glycoproteins - metabolism Molecular Sequence Data Neoplasm Proteins - isolation & purification Neoplasm Proteins - metabolism Recombinant Proteins - immunology Tumor Cells, Cultured |
| title | Production and purification of melanoma gp100 antigen and polyclonal antibodies |
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