Purification of a Soluble Hepatitis E Open Reading Frame 2-Derived Protein with Unique Antigenic Properties
The second open reading frame (ORF2) of hepatitis E virus (HEV) is predicted to encode a 73-kDa capsid protein (1). When full-length ORF2 was expressed in insect cells (Spodoptera frugiperda(Sf9)) using a recombinant baculovirus, two distinct HEV polypeptides were observed: a full-length insoluble 7...
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| Veröffentlicht in: | Protein expression and purification 1996-09, Vol.8 (2), p.262-270 |
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| creator | McAtee, C.Patrick Zhang, Yifan Yarbough, Patrice O. Bird, Thomas Fuerst, Thomas R. |
| description | The second open reading frame (ORF2) of hepatitis E virus (HEV) is predicted to encode a 73-kDa capsid protein (1). When full-length ORF2 was expressed in insect cells (Spodoptera frugiperda(Sf9)) using a recombinant baculovirus, two distinct HEV polypeptides were observed: a full-length insoluble 73-kDa protein, and a soluble 56.5-kDa protein. Following purification and sequence analysis, it was determined that the 56.5-kDa protein was derived from endoproteolytic cleavage site that was between the Thr and Ala residues located at amino acids 111 and 112 in the ORF2 sequence with the carboxy terminus corresponding to residue 636 of the ORF2 sequence. Comparative ELISA data using human acute-phase antisera demonstrated that the 56.5-kDa protein served as a highly reactive antigen in detecting anti-HEV antibodies. These data suggest that the 56.5-kDa protein may serve as a particularly useful antigen for both diagnostic and vaccine purposes. |
| doi_str_mv | 10.1006/prep.1996.0099 |
| format | Article |
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These data suggest that the 56.5-kDa protein may serve as a particularly useful antigen for both diagnostic and vaccine purposes.</description><identifier>ISSN: 1046-5928</identifier><identifier>EISSN: 1096-0279</identifier><identifier>DOI: 10.1006/prep.1996.0099</identifier><identifier>PMID: 8812876</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Antigens, Viral - immunology ; Baculoviridae - genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hepatitis E - diagnosis ; Hepatitis E virus - chemistry ; Hepatitis E virus - genetics ; Mass Spectrometry ; Molecular Sequence Data ; Open Reading Frames - genetics ; Peptide Mapping ; Peptides - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Sequence Analysis ; Spodoptera - genetics ; Trypsin - metabolism ; Viral Proteins - immunology ; Viral Proteins - isolation & purification</subject><ispartof>Protein expression and purification, 1996-09, Vol.8 (2), p.262-270</ispartof><rights>1996 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-99d4e5d2bb56db904200cf78824e27cdd9688ed47137164024f8e28e6ef303673</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/prep.1996.0099$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3549,27923,27924,45994</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8812876$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>McAtee, C.Patrick</creatorcontrib><creatorcontrib>Zhang, Yifan</creatorcontrib><creatorcontrib>Yarbough, Patrice O.</creatorcontrib><creatorcontrib>Bird, Thomas</creatorcontrib><creatorcontrib>Fuerst, Thomas R.</creatorcontrib><title>Purification of a Soluble Hepatitis E Open Reading Frame 2-Derived Protein with Unique Antigenic Properties</title><title>Protein expression and purification</title><addtitle>Protein Expr Purif</addtitle><description>The second open reading frame (ORF2) of hepatitis E virus (HEV) is predicted to encode a 73-kDa capsid protein (1). When full-length ORF2 was expressed in insect cells (Spodoptera frugiperda(Sf9)) using a recombinant baculovirus, two distinct HEV polypeptides were observed: a full-length insoluble 73-kDa protein, and a soluble 56.5-kDa protein. Following purification and sequence analysis, it was determined that the 56.5-kDa protein was derived from endoproteolytic cleavage site that was between the Thr and Ala residues located at amino acids 111 and 112 in the ORF2 sequence with the carboxy terminus corresponding to residue 636 of the ORF2 sequence. Comparative ELISA data using human acute-phase antisera demonstrated that the 56.5-kDa protein served as a highly reactive antigen in detecting anti-HEV antibodies. These data suggest that the 56.5-kDa protein may serve as a particularly useful antigen for both diagnostic and vaccine purposes.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antigens, Viral - immunology</subject><subject>Baculoviridae - genetics</subject><subject>Blotting, Western</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Gene Expression</subject><subject>Hepatitis E - diagnosis</subject><subject>Hepatitis E virus - chemistry</subject><subject>Hepatitis E virus - genetics</subject><subject>Mass Spectrometry</subject><subject>Molecular Sequence Data</subject><subject>Open Reading Frames - genetics</subject><subject>Peptide Mapping</subject><subject>Peptides - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Sequence Analysis</subject><subject>Spodoptera - genetics</subject><subject>Trypsin - metabolism</subject><subject>Viral Proteins - immunology</subject><subject>Viral Proteins - isolation & purification</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kEtLxTAQhYMovrfuhKzc9ZqkbZosxTcIio91aJOpjvamNUkV_70t9-LO1QxzzhxmPkKOOFtwxuTpEGBYcK3lgjGtN8guZ1pmTFR6c-4LmZVaqB2yF-M7Y5xLVm6TbaW4UJXcJR8PY8AWbZ2w97RvaU2f-m5sOqA3MEzThJFe0vsBPH2E2qF_pVehXgIV2QUE_AJHH0KfAD39xvRGXzx-jkDPfMJX8GhndYCQEOIB2WrrLsLhuu6Tl6vL5_Ob7O7--vb87C6zea5TprUroHSiaUrpGs0KwZhtK6VEAaKyzmmpFLii4nnFZcFE0SoQCiS0Octlle-Tk1XuEPrplpjMEqOFrqs99GM0lcrzUqliMi5WRhv6GAO0Zgi4rMOP4czMdM1M18x0zUx3WjheJ4_NEtyffY1z0tVKh-m9L4RgokXwFhwGsMm4Hv-L_gWpT4kI</recordid><startdate>19960901</startdate><enddate>19960901</enddate><creator>McAtee, C.Patrick</creator><creator>Zhang, Yifan</creator><creator>Yarbough, Patrice O.</creator><creator>Bird, Thomas</creator><creator>Fuerst, Thomas R.</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>19960901</creationdate><title>Purification of a Soluble Hepatitis E Open Reading Frame 2-Derived Protein with Unique Antigenic Properties</title><author>McAtee, C.Patrick ; Zhang, Yifan ; Yarbough, Patrice O. ; Bird, Thomas ; Fuerst, Thomas R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-99d4e5d2bb56db904200cf78824e27cdd9688ed47137164024f8e28e6ef303673</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antigens, Viral - immunology</topic><topic>Baculoviridae - genetics</topic><topic>Blotting, Western</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Gene Expression</topic><topic>Hepatitis E - diagnosis</topic><topic>Hepatitis E virus - chemistry</topic><topic>Hepatitis E virus - genetics</topic><topic>Mass Spectrometry</topic><topic>Molecular Sequence Data</topic><topic>Open Reading Frames - genetics</topic><topic>Peptide Mapping</topic><topic>Peptides - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Sequence Analysis</topic><topic>Spodoptera - genetics</topic><topic>Trypsin - metabolism</topic><topic>Viral Proteins - immunology</topic><topic>Viral Proteins - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McAtee, C.Patrick</creatorcontrib><creatorcontrib>Zhang, Yifan</creatorcontrib><creatorcontrib>Yarbough, Patrice O.</creatorcontrib><creatorcontrib>Bird, Thomas</creatorcontrib><creatorcontrib>Fuerst, Thomas R.</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>McAtee, C.Patrick</au><au>Zhang, Yifan</au><au>Yarbough, Patrice O.</au><au>Bird, Thomas</au><au>Fuerst, Thomas R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification of a Soluble Hepatitis E Open Reading Frame 2-Derived Protein with Unique Antigenic Properties</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>1996-09-01</date><risdate>1996</risdate><volume>8</volume><issue>2</issue><spage>262</spage><epage>270</epage><pages>262-270</pages><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>The second open reading frame (ORF2) of hepatitis E virus (HEV) is predicted to encode a 73-kDa capsid protein (1). When full-length ORF2 was expressed in insect cells (Spodoptera frugiperda(Sf9)) using a recombinant baculovirus, two distinct HEV polypeptides were observed: a full-length insoluble 73-kDa protein, and a soluble 56.5-kDa protein. Following purification and sequence analysis, it was determined that the 56.5-kDa protein was derived from endoproteolytic cleavage site that was between the Thr and Ala residues located at amino acids 111 and 112 in the ORF2 sequence with the carboxy terminus corresponding to residue 636 of the ORF2 sequence. Comparative ELISA data using human acute-phase antisera demonstrated that the 56.5-kDa protein served as a highly reactive antigen in detecting anti-HEV antibodies. These data suggest that the 56.5-kDa protein may serve as a particularly useful antigen for both diagnostic and vaccine purposes.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8812876</pmid><doi>10.1006/prep.1996.0099</doi><tpages>9</tpages></addata></record> |
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| identifier | ISSN: 1046-5928 |
| ispartof | Protein expression and purification, 1996-09, Vol.8 (2), p.262-270 |
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| language | eng |
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| source | Elsevier ScienceDirect Journals Complete - AutoHoldings; MEDLINE |
| subjects | Amino Acid Sequence Animals Antigens, Viral - immunology Baculoviridae - genetics Blotting, Western Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Gene Expression Hepatitis E - diagnosis Hepatitis E virus - chemistry Hepatitis E virus - genetics Mass Spectrometry Molecular Sequence Data Open Reading Frames - genetics Peptide Mapping Peptides - chemistry Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Sequence Analysis Spodoptera - genetics Trypsin - metabolism Viral Proteins - immunology Viral Proteins - isolation & purification |
| title | Purification of a Soluble Hepatitis E Open Reading Frame 2-Derived Protein with Unique Antigenic Properties |
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