One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell)

Growth hormone (GH) was purified from African catfish (Clarias gariepinus) pituitary extracts in a single step by use of immunoaffinity chromatography. A monoclonal antibody to chicken GH, which labels the catfish hypophyseal somatotropes in immunocytochemistry, was coupled to CNBr-activated Sepharo...

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Bibliographic Details
Main Authors: Berghman, LR, Lescroart, Olivier, Roelants, Ivo, Ollevier, Frans, Kuhn, ER, Verhaert, PD, De Loof, Arnold, Van Leuven, Freddy, Vandesande, Frans
Format: Article in Journal/Newspaper
Language:English
Published: Pergamon-elsevier science ltd 1996
Subjects:
african catfish
immunoaffinity purification
monoclonal antibody
growth hormone
physicochemical characterization
microheterogeneity
bioactivity
triiodothyronine enhancement
salmon oncorhynchus-kisutch
eel anguilla-anguilla
coho salmon
rainbow-trout
pituitary-hormones
plasma thyroxine
heterogeneity
gairdneri
triiodothyronine
identification
Anguilla anguilla
Online Access:https://lirias.kuleuven.be/handle/123456789/61069
http://gateway.newisiknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ScienceDirect&SrcApp=PRODUCT_NAME&KeyAID=0305049195020985&DestApp=ScienceDirect&SrcAppSID=APP_SID&SrcJTitle=WURS_TITLE
Description
Summary:Growth hormone (GH) was purified from African catfish (Clarias gariepinus) pituitary extracts in a single step by use of immunoaffinity chromatography. A monoclonal antibody to chicken GH, which labels the catfish hypophyseal somatotropes in immunocytochemistry, was coupled to CNBr-activated Sepharose, and crude alkaline pituitary extracts were run over the immunoadsorbent. Reversed-phase high-performance liquid chromatography analysis of the eluted material suggested heterogeneity, whereas silver staining upon SDS-polyacrylamide gel electrophoresis showed one single band with an estimated molecular weight between 22,000 and 23,000 Da. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the same preparation revealed the presence of several components with molecular weights ranging from 20,170 to 20,900 Da. The amino terminus of the protein was homogeneous, and the first 50 residues matched the proposed sequence of GH from two other siluran species (Ictalunus punctatus and Pangasius pangasius), except for one substitution at position 3. These data unequivocally confirm the identity of the purified molecule as suggested by immunochemical evidence. The bioactivity of the GH preparation was demonstrated by the short-term effect of GH on T-3 plasma levels in juvenile catfish. status: published

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