Immunoaffinity purification and partial characterization of sea bass (Dicentrarchus labrax) growth hormone.
Growth hormone (GH) was isolated from sea bass (Dicentrarchus labrax) pituitary extract by a simple one-step procedure involving immunoaffinity chromatography. A monoclonal antibody raised against chicken GH and found to immunostain very specifically the GH cells in the pituitary of the sea bass was coupled to CNBr-activated Sepharose 4B. Sea bass pituitary extracts were run on the affinity column, and the eluted material was analyzed on reversed-phase HPLC and found to consist of one single peak. The yield of purified hormone was 2.4 mg/g pituitary. Two monomeric forms (MW = 20,000 and 22,000 Da) of sea bass GH were identified by gel electrophoresis. Gel electrofocusing revealed apparent isoelectric points of 6.15, 6.50, and 6.95. Amino acid composition is consistent with other vertebrate GHs. The immunological relatedness was tested by immunoblotting using antisera raised against GH of different species. Polyclonal antisera raised against the isolated hormone exhibited a specific labeling of the GH cells in sea bass pituitary sections as well as of the immunoblotted purified GH.
Laboratory for Neuroendocrinology and Immunological Biotechnology, Zoological Institute, Leuven, Belgium.
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Berghman
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Vandesande
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D001492:Bass / Q000502:physiology*; D002645:Chickens; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes; D013006:Growth Hormone / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D015151:Immunoblotting; D008970:Molecular Weight; D010902:Pituitary Gland / Q000737:chemistry / Q000166:cytology; D011817:Rabbits
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D013006:Growth Hormone