The development of a noncompetitive enzyme-linked immunosorbent assay for oncorhynchid growth hormone using monoclonal antibodies.
Identifieur interne : 000264 ( Ncbi/Curation ); précédent : 000263; suivant : 000265The development of a noncompetitive enzyme-linked immunosorbent assay for oncorhynchid growth hormone using monoclonal antibodies.
Auteurs : K J Farbridge [Canada] ; J F LeatherlandSource :
- General and comparative endocrinology [ 0016-6480 ] ; 1991.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal, Chromatography, Affinity, Cross Reactions, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay (methods), Growth Hormone (analysis), Growth Hormone (immunology), Immunohistochemistry, Male, Pituitary Gland (chemistry), Pituitary Gland, Anterior (chemistry), Salmon (metabolism), Species Specificity, Trout (metabolism).
- MESH :
- chemical , analysis : Growth Hormone.
- chemical , immunology : Growth Hormone.
- chemical : Antibodies, Monoclonal.
- chemistry : Pituitary Gland, Pituitary Gland, Anterior.
- metabolism : Salmon, Trout.
- methods : Enzyme-Linked Immunosorbent Assay.
- Animals, Chromatography, Affinity, Cross Reactions, Electrophoresis, Polyacrylamide Gel, Immunohistochemistry, Male, Species Specificity.
Abstract
The development of a sensitive and specific two-site, or sandwich, noncompetitive enzyme-linked immunosorbent assay (ELISA) for oncorhynchid growth hormone (GH) using monoclonal antibodies (MCAs) is reported. The MCAs were generated by the fusion of myeloma cells with spleen cells from mice that had been immunized with chum salmon (Oncorhynchus keta) recombinant GH. The MCAs specifically recognized the GH-secreting acidophils in the proximal pars distalis of immature male rainbow trout (Oncorhynchus mykiss) pituitaries. Affinity chromatography using one of the MCAs isolated a single protein with a molecular weight of 22,500 from a rainbow trout pituitary extract. The ELISA recognized recombinant chum salmon GH and the affinity-purified protein but did not recognize chum salmon prolactin, gonadotropin I or II, nor several mammalian hormone preparations. The ELISA recognized GH in rainbow trout, coho salmon (Oncorhynchus kisutch), and chinook salmon (Oncorhynchus tshawytscha) pituitary extracts, but not in goldfish (Carassius auratus) extracts, and recognized GH in rainbow trout, coho salmon, lake sturgeon (Acipenser fulvescens), and bowfin (Amia calva) plasma, but not in goldfish, yellow bullhead (Ictalurus natalis), or lamprey (Petromyzon marinus) plasma. The sensitivity of the ELISA was less than 1.56 ng/ml and circulating levels of GH in the plasma of coho salmon and rainbow trout plasma were measured as 75 and 35 ng equivalents/ml, respectively.
PubMed: 1879674
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">The development of a noncompetitive enzyme-linked immunosorbent assay for oncorhynchid growth hormone using monoclonal antibodies.</title><author><name sortKey="Farbridge, K J" sort="Farbridge, K J" uniqKey="Farbridge K" first="K J" last="Farbridge">K J Farbridge</name><affiliation wicri:level="1"><nlm:affiliation>Department of Zoology, University of Guelph, Ontario, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Zoology, University of Guelph, Ontario</wicri:regionArea><wicri:noRegion>Ontario</wicri:noRegion></affiliation></author><author><name sortKey="Leatherland, J F" sort="Leatherland, J F" uniqKey="Leatherland J" first="J F" last="Leatherland">J F Leatherland</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1991">1991</date><idno type="RBID">pubmed:1879674</idno><idno type="pmid">1879674</idno><idno type="wicri:Area/PubMed/Corpus">000735</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000735</idno><idno type="wicri:Area/PubMed/Curation">000735</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000735</idno><idno type="wicri:Area/PubMed/Checkpoint">000735</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000735</idno><idno type="wicri:Area/Ncbi/Merge">000264</idno><idno type="wicri:Area/Ncbi/Curation">000264</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">The development of a noncompetitive enzyme-linked immunosorbent assay for oncorhynchid growth hormone using monoclonal antibodies.</title><author><name sortKey="Farbridge, K J" sort="Farbridge, K J" uniqKey="Farbridge K" first="K J" last="Farbridge">K J Farbridge</name><affiliation wicri:level="1"><nlm:affiliation>Department of Zoology, University of Guelph, Ontario, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Zoology, University of Guelph, Ontario</wicri:regionArea><wicri:noRegion>Ontario</wicri:noRegion></affiliation></author><author><name sortKey="Leatherland, J F" sort="Leatherland, J F" uniqKey="Leatherland J" first="J F" last="Leatherland">J F Leatherland</name></author></analytic><series><title level="j">General and comparative endocrinology</title><idno type="ISSN">0016-6480</idno><imprint><date when="1991" type="published">1991</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal</term><term>Chromatography, Affinity</term><term>Cross Reactions</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Growth Hormone (analysis)</term><term>Growth Hormone (immunology)</term><term>Immunohistochemistry</term><term>Male</term><term>Pituitary Gland (chemistry)</term><term>Pituitary Gland, Anterior (chemistry)</term><term>Salmon (metabolism)</term><term>Species Specificity</term><term>Trout (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Growth Hormone</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Growth Hormone</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antibodies, Monoclonal</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Pituitary Gland</term><term>Pituitary Gland, Anterior</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Salmon</term><term>Trout</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chromatography, Affinity</term><term>Cross Reactions</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Immunohistochemistry</term><term>Male</term><term>Species Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The development of a sensitive and specific two-site, or sandwich, noncompetitive enzyme-linked immunosorbent assay (ELISA) for oncorhynchid growth hormone (GH) using monoclonal antibodies (MCAs) is reported. The MCAs were generated by the fusion of myeloma cells with spleen cells from mice that had been immunized with chum salmon (Oncorhynchus keta) recombinant GH. The MCAs specifically recognized the GH-secreting acidophils in the proximal pars distalis of immature male rainbow trout (Oncorhynchus mykiss) pituitaries. Affinity chromatography using one of the MCAs isolated a single protein with a molecular weight of 22,500 from a rainbow trout pituitary extract. The ELISA recognized recombinant chum salmon GH and the affinity-purified protein but did not recognize chum salmon prolactin, gonadotropin I or II, nor several mammalian hormone preparations. The ELISA recognized GH in rainbow trout, coho salmon (Oncorhynchus kisutch), and chinook salmon (Oncorhynchus tshawytscha) pituitary extracts, but not in goldfish (Carassius auratus) extracts, and recognized GH in rainbow trout, coho salmon, lake sturgeon (Acipenser fulvescens), and bowfin (Amia calva) plasma, but not in goldfish, yellow bullhead (Ictalurus natalis), or lamprey (Petromyzon marinus) plasma. The sensitivity of the ELISA was less than 1.56 ng/ml and circulating levels of GH in the plasma of coho salmon and rainbow trout plasma were measured as 75 and 35 ng equivalents/ml, respectively.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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