One-step immunoaffinity purification and partial characterisation of a growth hormone from the pituitary gland of the African catfish, Clarias gariepinus (Burchell)
Lescroart, O.; Berghman, L.R.; Roelants, I.; Ollevier, F.P.; Kuhn, E.; Van Leuven, F.; Vandesande, F. (1993). One-step immunoaffinity purification and partial characterisation of a growth hormone from the pituitary gland of the African catfish, Clarias gariepinus (Burchell), in: Seventh forum for applied biotechnology, PAND, Gent 30 September - October 1993, abstracts. pp. 57
In: (1993). Seventh forum for applied biotechnology, PAND, Gent 30 September - October 1993, abstracts. RUG: Gent. 150 pp., more
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Document type: Conference paper
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| Authors | | Top |
- Lescroart, O.
- Berghman, L.R.
- Roelants, I.
- Ollevier, F.P., more
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- Kuhn, E.
- Van Leuven, F.
- Vandesande, F.
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| Abstract |
Growth hormone (GH) wa8 purified from African catfish (Clarias gariepinus) pituitary extracts in a single step, using immuno-affinity chromatography. An anti-chicken GH monoclonal antibody, specifically cross-reacting with the African catfish hypophyseal somatotropes, was coupled to CNBr-activated Sepharose (5mg IgG/ml gel). Crude pituitary extracts were run as such over the immunoadsorbent. Although reversed-phase HPLC analysis of the eluted material showed two partially matching peaks, subsequent SDS-PAGE showed one single band upon silver staining. The apparent molecular weight of the purified molecule (as assessed by SDS-PAGE) was approximately 23,000 Da. The amino terminal end of the protein was homogenous and with the exception of 1 substitution at position 3, the first 30 residus matched the proposed protein sequence for GH of the channel catfish (Ictalurus punctatus). This finding confirmed unequivocally the identity of the purified molecule that was formerly suggested by immunochemical evidence. |
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