Production and characterization of guinea pig IL-5 in baculovirus-infected insect cells
M. Mansour, M. Karmilowicz, S. J. Hawrylik, B. Nalcerio, J. Angilly, M. J. Conklyn, C. M. Lilly, J. M. Drazen, S. E. Lee, D. D. Auperin, J. R. De Wet, V. L. Cohan, H. J. Showell and D. E. Danley Department of Molecular Sciences, Pfizer Central Research Division, Groton, Connecticut 06340, USA. To st...
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| Veröffentlicht in: | American journal of physiology. Lung cellular and molecular physiology 1996-06, Vol.270 (6), p.1002-L1007 |
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| container_title | American journal of physiology. Lung cellular and molecular physiology |
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| creator | Mansour, M Karmilowicz, M Hawrylik, S. J Nalcerio, B Angilly, J Conklyn, M. J Lilly, C. M Drazen, J. M Lee, S. E Auperin, D. D De Wet, J. R Cohan, V. L Showell, H. J Danley, D. E |
| description | M. Mansour, M. Karmilowicz, S. J. Hawrylik, B. Nalcerio, J. Angilly, M. J. Conklyn, C. M. Lilly, J. M. Drazen, S. E. Lee, D. D. Auperin, J. R. De Wet, V. L. Cohan, H. J. Showell and D. E. Danley
Department of Molecular Sciences, Pfizer Central Research Division, Groton, Connecticut 06340, USA.
To study the role interleukin (IL)-5 may play in altering airway function
in asthma, we have produced recombinant protein for exogenous
administration to guinea pigs. The guinea pig IL-5 (gpIL-5) cDNA was cloned
by polymerase chain reaction (PCR) amplification of guinea pig spleen RNA
and expressed as a secretion product from recombinant baculovirus-infected
Sf9 insect cell cultures. The protein was purified to homogeneity by a
four-step procedure that included immunoaffinity chromatography using
polyclonal antipeptide antibodies against a region of the mature secreted
cytokine. The cytokine was properly processed after the signal sequence by
the Sf9 cells, was glycosylated with terminal mannose-containing
oligosaccharide, and had proper disulfide-linked dimer structure as
determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The purified preparation was active in vitro and in vivo as determined by
its ability to prime human basophils to release leukotriene C4 in the
presence of C5a and to induce airway eosinophilia in naive guinea pigs. |
| doi_str_mv | 10.1152/ajplung.1996.270.6.l1002 |
| format | Article |
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Department of Molecular Sciences, Pfizer Central Research Division, Groton, Connecticut 06340, USA.
To study the role interleukin (IL)-5 may play in altering airway function
in asthma, we have produced recombinant protein for exogenous
administration to guinea pigs. The guinea pig IL-5 (gpIL-5) cDNA was cloned
by polymerase chain reaction (PCR) amplification of guinea pig spleen RNA
and expressed as a secretion product from recombinant baculovirus-infected
Sf9 insect cell cultures. The protein was purified to homogeneity by a
four-step procedure that included immunoaffinity chromatography using
polyclonal antipeptide antibodies against a region of the mature secreted
cytokine. The cytokine was properly processed after the signal sequence by
the Sf9 cells, was glycosylated with terminal mannose-containing
oligosaccharide, and had proper disulfide-linked dimer structure as
determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The purified preparation was active in vitro and in vivo as determined by
its ability to prime human basophils to release leukotriene C4 in the
presence of C5a and to induce airway eosinophilia in naive guinea pigs.</description><identifier>ISSN: 1040-0605</identifier><identifier>ISSN: 0002-9513</identifier><identifier>EISSN: 1522-1504</identifier><identifier>DOI: 10.1152/ajplung.1996.270.6.l1002</identifier><identifier>PMID: 8764226</identifier><language>eng</language><publisher>United States</publisher><subject>Animals ; Baculoviridae ; Base Sequence ; Cell Line ; Cloning, Molecular ; Guinea Pigs ; Humans ; Insecta - virology ; Interleukin-5 - genetics ; Interleukin-5 - isolation & purification ; Interleukin-5 - physiology ; Male ; Mice ; Molecular Sequence Data ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Sequence Homology, Amino Acid</subject><ispartof>American journal of physiology. Lung cellular and molecular physiology, 1996-06, Vol.270 (6), p.1002-L1007</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8764226$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mansour, M</creatorcontrib><creatorcontrib>Karmilowicz, M</creatorcontrib><creatorcontrib>Hawrylik, S. J</creatorcontrib><creatorcontrib>Nalcerio, B</creatorcontrib><creatorcontrib>Angilly, J</creatorcontrib><creatorcontrib>Conklyn, M. J</creatorcontrib><creatorcontrib>Lilly, C. M</creatorcontrib><creatorcontrib>Drazen, J. M</creatorcontrib><creatorcontrib>Lee, S. E</creatorcontrib><creatorcontrib>Auperin, D. D</creatorcontrib><creatorcontrib>De Wet, J. R</creatorcontrib><creatorcontrib>Cohan, V. L</creatorcontrib><creatorcontrib>Showell, H. J</creatorcontrib><creatorcontrib>Danley, D. E</creatorcontrib><title>Production and characterization of guinea pig IL-5 in baculovirus-infected insect cells</title><title>American journal of physiology. Lung cellular and molecular physiology</title><addtitle>Am J Physiol</addtitle><description>M. Mansour, M. Karmilowicz, S. J. Hawrylik, B. Nalcerio, J. Angilly, M. J. Conklyn, C. M. Lilly, J. M. Drazen, S. E. Lee, D. D. Auperin, J. R. De Wet, V. L. Cohan, H. J. Showell and D. E. Danley
Department of Molecular Sciences, Pfizer Central Research Division, Groton, Connecticut 06340, USA.
To study the role interleukin (IL)-5 may play in altering airway function
in asthma, we have produced recombinant protein for exogenous
administration to guinea pigs. The guinea pig IL-5 (gpIL-5) cDNA was cloned
by polymerase chain reaction (PCR) amplification of guinea pig spleen RNA
and expressed as a secretion product from recombinant baculovirus-infected
Sf9 insect cell cultures. The protein was purified to homogeneity by a
four-step procedure that included immunoaffinity chromatography using
polyclonal antipeptide antibodies against a region of the mature secreted
cytokine. The cytokine was properly processed after the signal sequence by
the Sf9 cells, was glycosylated with terminal mannose-containing
oligosaccharide, and had proper disulfide-linked dimer structure as
determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The purified preparation was active in vitro and in vivo as determined by
its ability to prime human basophils to release leukotriene C4 in the
presence of C5a and to induce airway eosinophilia in naive guinea pigs.</description><subject>Animals</subject><subject>Baculoviridae</subject><subject>Base Sequence</subject><subject>Cell Line</subject><subject>Cloning, Molecular</subject><subject>Guinea Pigs</subject><subject>Humans</subject><subject>Insecta - virology</subject><subject>Interleukin-5 - genetics</subject><subject>Interleukin-5 - isolation & purification</subject><subject>Interleukin-5 - physiology</subject><subject>Male</subject><subject>Mice</subject><subject>Molecular Sequence Data</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Sequence Homology, Amino Acid</subject><issn>1040-0605</issn><issn>0002-9513</issn><issn>1522-1504</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkMtOwzAQRS0EgvL4BCSv2CWMHTuul6jiUakSLEAsLcdxUldpHOwGVL4el1aI1Yzmzr0zOghhAjkhnN7q1dCNfZsTKcucCsjLvCMA9AhNkkwzwoEdpx4YZFACP0PnMa4AgAOUp-h0KkpGaTlB7y_B16PZON9j3dfYLHXQZmOD-9a_Q9_gdnS91XhwLZ4vMo5djyttxs5_ujDGzPWNTY46zWNqsLFdFy_RSaO7aK8O9QK9Pdy_zp6yxfPjfHa3yAyVbJMxM5WFqSUtNFTCGA2NqASXtqqFIZIaK6hoaKkJ1cxwKFghaGEaMJIVtWyKC3Szzx2C_xht3Ki1i7sPdG_9GJWYEs445Wlxul80wccYbKOG4NY6bBUBtWOqDkzVjqlKTFWpFjumyXp9uDFWa1v_GQ8Qk57v9aVrl18uWDUst9H5zrfbv9T_gT8P9YYq</recordid><startdate>19960601</startdate><enddate>19960601</enddate><creator>Mansour, M</creator><creator>Karmilowicz, M</creator><creator>Hawrylik, S. 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E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c294t-4c893cd923a0b7cca0f7b759ebd7c192ce727f26a12a4c50343723cf0c943d9f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Baculoviridae</topic><topic>Base Sequence</topic><topic>Cell Line</topic><topic>Cloning, Molecular</topic><topic>Guinea Pigs</topic><topic>Humans</topic><topic>Insecta - virology</topic><topic>Interleukin-5 - genetics</topic><topic>Interleukin-5 - isolation & purification</topic><topic>Interleukin-5 - physiology</topic><topic>Male</topic><topic>Mice</topic><topic>Molecular Sequence Data</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Sequence Homology, Amino Acid</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mansour, M</creatorcontrib><creatorcontrib>Karmilowicz, M</creatorcontrib><creatorcontrib>Hawrylik, S. J</creatorcontrib><creatorcontrib>Nalcerio, B</creatorcontrib><creatorcontrib>Angilly, J</creatorcontrib><creatorcontrib>Conklyn, M. J</creatorcontrib><creatorcontrib>Lilly, C. M</creatorcontrib><creatorcontrib>Drazen, J. M</creatorcontrib><creatorcontrib>Lee, S. E</creatorcontrib><creatorcontrib>Auperin, D. D</creatorcontrib><creatorcontrib>De Wet, J. R</creatorcontrib><creatorcontrib>Cohan, V. L</creatorcontrib><creatorcontrib>Showell, H. J</creatorcontrib><creatorcontrib>Danley, D. E</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>American journal of physiology. Lung cellular and molecular physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mansour, M</au><au>Karmilowicz, M</au><au>Hawrylik, S. J</au><au>Nalcerio, B</au><au>Angilly, J</au><au>Conklyn, M. J</au><au>Lilly, C. M</au><au>Drazen, J. M</au><au>Lee, S. E</au><au>Auperin, D. D</au><au>De Wet, J. R</au><au>Cohan, V. L</au><au>Showell, H. J</au><au>Danley, D. E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production and characterization of guinea pig IL-5 in baculovirus-infected insect cells</atitle><jtitle>American journal of physiology. Lung cellular and molecular physiology</jtitle><addtitle>Am J Physiol</addtitle><date>1996-06-01</date><risdate>1996</risdate><volume>270</volume><issue>6</issue><spage>1002</spage><epage>L1007</epage><pages>1002-L1007</pages><issn>1040-0605</issn><issn>0002-9513</issn><eissn>1522-1504</eissn><abstract>M. Mansour, M. Karmilowicz, S. J. Hawrylik, B. Nalcerio, J. Angilly, M. J. Conklyn, C. M. Lilly, J. M. Drazen, S. E. Lee, D. D. Auperin, J. R. De Wet, V. L. Cohan, H. J. Showell and D. E. Danley
Department of Molecular Sciences, Pfizer Central Research Division, Groton, Connecticut 06340, USA.
To study the role interleukin (IL)-5 may play in altering airway function
in asthma, we have produced recombinant protein for exogenous
administration to guinea pigs. The guinea pig IL-5 (gpIL-5) cDNA was cloned
by polymerase chain reaction (PCR) amplification of guinea pig spleen RNA
and expressed as a secretion product from recombinant baculovirus-infected
Sf9 insect cell cultures. The protein was purified to homogeneity by a
four-step procedure that included immunoaffinity chromatography using
polyclonal antipeptide antibodies against a region of the mature secreted
cytokine. The cytokine was properly processed after the signal sequence by
the Sf9 cells, was glycosylated with terminal mannose-containing
oligosaccharide, and had proper disulfide-linked dimer structure as
determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The purified preparation was active in vitro and in vivo as determined by
its ability to prime human basophils to release leukotriene C4 in the
presence of C5a and to induce airway eosinophilia in naive guinea pigs.</abstract><cop>United States</cop><pmid>8764226</pmid><doi>10.1152/ajplung.1996.270.6.l1002</doi></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1040-0605 |
| ispartof | American journal of physiology. Lung cellular and molecular physiology, 1996-06, Vol.270 (6), p.1002-L1007 |
| issn | 1040-0605 0002-9513 1522-1504 |
| language | eng |
| recordid | cdi_highwire_physiology_ajplung_270_6_L1002 |
| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Animals Baculoviridae Base Sequence Cell Line Cloning, Molecular Guinea Pigs Humans Insecta - virology Interleukin-5 - genetics Interleukin-5 - isolation & purification Interleukin-5 - physiology Male Mice Molecular Sequence Data Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Sequence Homology, Amino Acid |
| title | Production and characterization of guinea pig IL-5 in baculovirus-infected insect cells |
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