Production and characterization of an antiserum against recombinant porcine follicle stimulating hormone
This study aimed to develop an antiserum specific to follicle stimulating hormone (FSH), for a wide range of species, free from cross-reactivity against luteinizing hormone. An antiserum against recombinant porcine FSH (rec-pFSH) expressed in insect cells was raised in a rabbit and its specificity w...
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| Published in | Journal of Reproduction and Development Vol. 48; no. 2; pp. 131 - 136 |
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| Main Authors | , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT
2002
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0916-8818 1348-4400 |
| DOI | 10.1262/jrd.48.131 |
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| Abstract | This study aimed to develop an antiserum specific to follicle stimulating hormone (FSH), for a wide range of species, free from cross-reactivity against luteinizing hormone. An antiserum against recombinant porcine FSH (rec-pFSH) expressed in insect cells was raised in a rabbit and its specificity was characterized by time-resolved fluoroimmunoassay (TR-FIA) and immunohistochemistry. The titer of the anti-rec-pFSH antiserum was about 1:5,000 for use in TR-FIA. The inhibition curve of TR-FIA with rec-pFSH paralleled that of pituitary-derived FSH, indicating that the antigenicities of the two FSH preparations do not differ significantly. The anti-rec-pFSH antiserum showed low reactivities with FSHs prepared from sheep and rat, suggesting it was species- and hormone-specific, contrary to our intention. The TR-FIA showed that this antiserum did not react with pituitary extracts from mammals other than the pig. However, immunohistochemistry demonstrated that this antiserum reacted with the specific cells not only of the porcine anterior pituitary but also of the cow, goat, and horse. The antiserum also immunostained pituitary cells of the cow, goat and horse. Therefore, this anti-rec-pFSH antiserum will be of limited use in studies involving competitive immunoreactions due to its species-specific reactivity, but it should be useful for non-competitive techniques, such as immunohistochemistry. |
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| AbstractList | This study aimed to develop an antiserum specific to follicle stimulating hormone (FSH), for a wide range of species, free from cross-reactivity against luteinizing hormone. An antiserum against recombinant porcine FSH (rec-pFSH) expressed in insect cells was raised in a rabbit and its specificity was characterized by time-resolved fluoroimmunoassay (TR-FIA) and immunohistochemistry. The titer of the anti-rec-pFSH antiserum was about 1:5,000 for use in TR-FIA. The inhibition curve of TR-FIA with rec-pFSH paralleled that of pituitary-derived FSH, indicating that the antigenicities of the two FSH preparations do not differ significantly. The anti-rec-pFSH antiserum showed low reactivities with FSHs prepared from sheep and rat, suggesting it was species- and hormone-specific, contrary to our intention. The TR-FIA showed that this antiserum did not react with pituitary extracts from mammals other than the pig. However, immunohistochemistry demonstrated that this antiserum reacted with the specific cells not only of the porcine anterior pituitary but also of the cow, goat, and horse. The antiserum also immunostained pituitary cells of the cow, goat and horse. Therefore, this anti-rec-pFSH antiserum will be of limited use in studies involving competitive immunoreactions due to its species-specific reactivity, but it should be useful for non-competitive techniques, such as immunohistochemistry. |
| Author | Kato, T Sakai, T Watanabe, M Ueda, S Sato, I Inoue, K Ogawa, S Tomizawa, K. (Gunma Univ., Maebashi (Japan)) Ihara, T Kikuchi, M Kato, Y Hattori, M Mori, J |
| Author_xml | – sequence: 1 fullname: Tomizawa, K. (Gunma Univ., Maebashi (Japan)) – sequence: 2 fullname: Ogawa, S – sequence: 3 fullname: Kato, T – sequence: 4 fullname: Watanabe, M – sequence: 5 fullname: Sato, I – sequence: 6 fullname: Ihara, T – sequence: 7 fullname: Sakai, T – sequence: 8 fullname: Hattori, M – sequence: 9 fullname: Inoue, K – sequence: 10 fullname: Ueda, S – sequence: 11 fullname: Kikuchi, M – sequence: 12 fullname: Mori, J – sequence: 13 fullname: Kato, Y |
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| Cites_doi | 10.1016/S0093-691X(97)00007-1 10.1677/jme.0.0200055 10.1677/jme.0.0120071 10.4049/jimmunol.127.3.1244 10.1016/S0021-9258(18)83656-0 10.1507/endocrj1954.34.755 10.1016/S0034-5288(98)90110-2 10.1267/ahc.21.221 |
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| References | 9. Iwasawa A, Tomizawa K, Kato T, Wakabayashi K, Kato Y . Time-resolved fluoroimmunoassay (TR-FIA) of gonadotropins. Exp Clin Endocrinol 1994 ; 102 : 40 -44. 1. Keene JL, Matzuk MM, Otani T, Fauser BCJM, Galway AB, Hsueh AJW, Boime I . Expression of biologically active human follitropin in Chinese hamster ovary cells. J Biol Chem 1989 ; 264 : 4769 -4775. 10. Hunter R, Bennett B . The adjuvant activity of nonionic block polymer surfactants. II. Antibody formation and inflammation related to the structure of triblock and octablock copolymers. J Immunol 1981 ; 133 : 167 -3175. 4. Kato Y, Sato I, Ihara T, Tomizawa K, Mori J, Geshi M, Nagai T, Okuda K, Kato T, Ueda S . Expression of biologically active porcine follicle stimulating hormone in insect cells bearing a baculovirus vector and its purification. J Mol Endocrinol 1998 ; 20 : 55 -65. 11. Hunter R, Strickland F, Kezdy F . The adjuvant activity of nonionic block copolymer surfactants. I. The role of hydrophile-lipophile balance. J Immunol 1981 ; 127 : 1244 -1250. 3. Van Wezenbeek P, Draaijer J, Meel FV, Olijve W . Recombinant follicle stimulating hormone. I. Construction, selection and characterization of a cell line. In: Crommelin DJA, Schellekens H (eds.), Clone to Clinic Development in Biotherapy. , Dordrecht: Kluwer Academic Publisher; 1990 , Vol. 1 : 245 -251. 2. Mountford PS, Brandon MR, Adams TE . Expression and characterization of biologically active ovine FSH from mammalian cell lines. J Mol Endocrinol 1995 ; 12 : 71 -83. 6. Inaba T, Mori J, Ohmura M, Tomizawa K, Kato Y, Ihara T, Sato I, Ueda S . Baculovirus-insect cell production of bioactive porcine FSH. Theriogenology 1997 ; 47 : 491 -499. 12. Wakabayashi K, Tanaka S . Assessment of specificity of antisera for immunohistochemistry. Acta Histochem Cytochem 1988 ; 21 : 221 -229. 7. Ishikawa J, Fuse Y, Wakabayashi K . Choice of extraction procedure for estimation of anterior pituitary hormone content. Endocrino J 1987 ; 34 : 755 -767. 8. Goudie RB, Horne CHW, Wilkinson PC . A simple method for producing antibody specific to a single selected diffusible antigen. Lancet 1966 ; 2 : 1224 -1226. 5. Inaba T, Mori J, Ohmura M, Tani H, Kato Y, Tomizawa K, Kato T, Ihara T, Sato I, Ueda S . Recombinant porcine follicle stimulating hormone produced in baculovirus-insect cells induces rat ovulation in vivo and gene expression of tissue plasminogen activator in vitro . Res Vet Sci 1998 ; 64 : 25 -29. Ishikawa J, Fuse Y, Wakabayashi K (7) 1987; 34 Hunter R, Strickland F, Kezdy F (11) 1981; 127 (4) 1998; 20 5 6 Iwasawa A, Tomizawa K, Kato T, Waka (9) 1994; 102 Mountford PS, Brandon MR, Adams TE (2) 1995; 12 Goudie RB, Horne CHW, Wilkinson PC (8) 1966; 2 WAKABAYASHI K (12) 1988; 21 Keene JL, Matzuk MM, Otani T, Fause (1) 1989; 264 Van Wezenbeek P, Draaijer J, Meel F (3) 1990; 1 Hunter R, Bennett B (10) 1981; 133 |
| References_xml | – reference: 3. Van Wezenbeek P, Draaijer J, Meel FV, Olijve W . Recombinant follicle stimulating hormone. I. Construction, selection and characterization of a cell line. In: Crommelin DJA, Schellekens H (eds.), Clone to Clinic Development in Biotherapy. , Dordrecht: Kluwer Academic Publisher; 1990 , Vol. 1 : 245 -251. – reference: 12. Wakabayashi K, Tanaka S . Assessment of specificity of antisera for immunohistochemistry. Acta Histochem Cytochem 1988 ; 21 : 221 -229. – reference: 1. Keene JL, Matzuk MM, Otani T, Fauser BCJM, Galway AB, Hsueh AJW, Boime I . Expression of biologically active human follitropin in Chinese hamster ovary cells. J Biol Chem 1989 ; 264 : 4769 -4775. – reference: 4. Kato Y, Sato I, Ihara T, Tomizawa K, Mori J, Geshi M, Nagai T, Okuda K, Kato T, Ueda S . Expression of biologically active porcine follicle stimulating hormone in insect cells bearing a baculovirus vector and its purification. J Mol Endocrinol 1998 ; 20 : 55 -65. – reference: 6. Inaba T, Mori J, Ohmura M, Tomizawa K, Kato Y, Ihara T, Sato I, Ueda S . Baculovirus-insect cell production of bioactive porcine FSH. Theriogenology 1997 ; 47 : 491 -499. – reference: 8. Goudie RB, Horne CHW, Wilkinson PC . A simple method for producing antibody specific to a single selected diffusible antigen. Lancet 1966 ; 2 : 1224 -1226. – reference: 2. Mountford PS, Brandon MR, Adams TE . Expression and characterization of biologically active ovine FSH from mammalian cell lines. J Mol Endocrinol 1995 ; 12 : 71 -83. – reference: 5. Inaba T, Mori J, Ohmura M, Tani H, Kato Y, Tomizawa K, Kato T, Ihara T, Sato I, Ueda S . Recombinant porcine follicle stimulating hormone produced in baculovirus-insect cells induces rat ovulation in vivo and gene expression of tissue plasminogen activator in vitro . Res Vet Sci 1998 ; 64 : 25 -29. – reference: 10. Hunter R, Bennett B . The adjuvant activity of nonionic block polymer surfactants. II. Antibody formation and inflammation related to the structure of triblock and octablock copolymers. J Immunol 1981 ; 133 : 167 -3175. – reference: 7. Ishikawa J, Fuse Y, Wakabayashi K . Choice of extraction procedure for estimation of anterior pituitary hormone content. Endocrino J 1987 ; 34 : 755 -767. – reference: 9. Iwasawa A, Tomizawa K, Kato T, Wakabayashi K, Kato Y . Time-resolved fluoroimmunoassay (TR-FIA) of gonadotropins. Exp Clin Endocrinol 1994 ; 102 : 40 -44. – reference: 11. Hunter R, Strickland F, Kezdy F . The adjuvant activity of nonionic block copolymer surfactants. I. The role of hydrophile-lipophile balance. J Immunol 1981 ; 127 : 1244 -1250. – ident: 6 doi: 10.1016/S0093-691X(97)00007-1 – volume: 102 start-page: 40 issn: 0232-7384 year: 1994 ident: 9 publication-title: Exp Clin Endocrinol – volume: 20 start-page: 55 issn: 0952-5041 issue: 1 year: 1998 ident: 4 publication-title: J Mol Endocrinol doi: 10.1677/jme.0.0200055 – volume: 12 start-page: 71 issn: 0952-5041 year: 1995 ident: 2 publication-title: J Mol Endocrinol doi: 10.1677/jme.0.0120071 – volume: 1 start-page: 245 year: 1990 ident: 3 – volume: 127 start-page: 1244 issn: 0022-1767 year: 1981 ident: 11 publication-title: J Immunol doi: 10.4049/jimmunol.127.3.1244 – volume: 264 start-page: 4769 issn: 0021-9258 issue: 9 year: 1989 ident: 1 publication-title: J Biol Chem doi: 10.1016/S0021-9258(18)83656-0 – volume: 34 start-page: 755 year: 1987 ident: 7 publication-title: Endocrino J doi: 10.1507/endocrj1954.34.755 – ident: 5 doi: 10.1016/S0034-5288(98)90110-2 – volume: 2 start-page: 1224 issn: 0140-6736 year: 1966 ident: 8 publication-title: Lancet – volume: 21 start-page: 221 issn: 0044-5991 issue: 3 year: 1988 ident: 12 publication-title: Acta Histochem Cytochem doi: 10.1267/ahc.21.221 – volume: 133 start-page: 167 issn: 0022-1767 year: 1981 ident: 10 publication-title: J Immunol |
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| SubjectTerms | Antibody FSH Immunohistochemistry Pig Pituitary hormone PITUITARY HORMONES Recombinant RECOMBINANT DNA SWINE |
| Title | Production and characterization of an antiserum against recombinant porcine follicle stimulating hormone |
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