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...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Pergamon-elsevier science ltd
1996
|
Subjects: | |
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 |
id |
ftunivleuven:oai:lirias.kuleuven.be:123456789/61069 |
---|---|
record_format |
openpolar |
spelling |
ftunivleuven:oai:lirias.kuleuven.be:123456789/61069 2023-05-15T13:27:54+02:00 One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) Berghman, LR Lescroart, Olivier Roelants, Ivo Ollevier, Frans Kuhn, ER Verhaert, PD De Loof, Arnold Van Leuven, Freddy Vandesande, Frans 1996-04 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 en eng Pergamon-elsevier science ltd Comparative biochemistry and physiology b-biochemistry & molecular biology vol:113 issue:4 pages:773-780 https://lirias.kuleuven.be/handle/123456789/61069 0305-0491 http://gateway.newisiknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ScienceDirect&SrcApp=PRODUCT_NAME&KeyAID=0305049195020985&DestApp=ScienceDirect&SrcAppSID=APP_SID&SrcJTitle=WURS_TITLE 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 Description (Metadata) only IT article 1996 ftunivleuven 2015-12-22T15:09:57Z 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 Article in Journal/Newspaper Anguilla anguilla KU Leuven: Lirias |
institution |
Open Polar |
collection |
KU Leuven: Lirias |
op_collection_id |
ftunivleuven |
language |
English |
topic |
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 |
spellingShingle |
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 Berghman, LR Lescroart, Olivier Roelants, Ivo Ollevier, Frans Kuhn, ER Verhaert, PD De Loof, Arnold Van Leuven, Freddy Vandesande, Frans One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
topic_facet |
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 |
description |
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 |
format |
Article in Journal/Newspaper |
author |
Berghman, LR Lescroart, Olivier Roelants, Ivo Ollevier, Frans Kuhn, ER Verhaert, PD De Loof, Arnold Van Leuven, Freddy Vandesande, Frans |
author_facet |
Berghman, LR Lescroart, Olivier Roelants, Ivo Ollevier, Frans Kuhn, ER Verhaert, PD De Loof, Arnold Van Leuven, Freddy Vandesande, Frans |
author_sort |
Berghman, LR |
title |
One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
title_short |
One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
title_full |
One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
title_fullStr |
One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
title_full_unstemmed |
One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell) |
title_sort |
one-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the african catfish, clarias gariepinus (burchell) |
publisher |
Pergamon-elsevier science ltd |
publishDate |
1996 |
url |
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 |
genre |
Anguilla anguilla |
genre_facet |
Anguilla anguilla |
op_relation |
Comparative biochemistry and physiology b-biochemistry & molecular biology vol:113 issue:4 pages:773-780 https://lirias.kuleuven.be/handle/123456789/61069 0305-0491 http://gateway.newisiknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ScienceDirect&SrcApp=PRODUCT_NAME&KeyAID=0305049195020985&DestApp=ScienceDirect&SrcAppSID=APP_SID&SrcJTitle=WURS_TITLE |
_version_ |
1766401035719409664 |