Lab-scale Preparation of Recombinant Human Insulin-like Growth Factor-1 in
ELISA
Escherichia coli
Western blot
hIGF-1
human lung cell line
proliferation assay
recombinant expression
Journal
Recent patents on biotechnology
ISSN: 2212-4012
Titre abrégé: Recent Pat Biotechnol
Pays: United Arab Emirates
ID NLM: 101309942
Informations de publication
Date de publication:
03 08 2022
03 08 2022
Historique:
received:
11
11
2021
revised:
30
01
2022
accepted:
03
03
2022
pubmed:
15
4
2022
medline:
19
8
2022
entrez:
14
4
2022
Statut:
ppublish
Résumé
Insulin-like growth factor-1 (IGF-1) is structurally similar to insulin and acts as an endocrine hormone secreted by the liver. Production of recombinant human IGF-1 (rhIGF-1) in hIGF-1 gene cloned into pBSK (+) simple vector was transformed into TOP 10 chemically competent cells of rhIGF-1 was purified from the HiTrap-ANX column at a concentration of 300 μg/ml. Western blot showed a single 7.6 kDa band obtained in the induced Rosetta (DE3) pLYsS. ELISA confirmed the molecular identity of the rhIGF-1 epitope, the concentration of purified rhIGF-1 obtained from the ELISA standard curve using rhIGF-1 reference protein as a standard was 300 μg/ml, and activity on WI-38 cells was 2604.17I U/mg. Biologically active native rhIGF-1 protein was successfully expressed. Patents related to the preparation of IGF-1 were mentioned along the text.
Sections du résumé
BACKGROUND
Insulin-like growth factor-1 (IGF-1) is structurally similar to insulin and acts as an endocrine hormone secreted by the liver.
OBJECTIVE
Production of recombinant human IGF-1 (rhIGF-1) in
METHODS
hIGF-1 gene cloned into pBSK (+) simple vector was transformed into TOP 10 chemically competent cells of
RESULTS
rhIGF-1 was purified from the HiTrap-ANX column at a concentration of 300 μg/ml. Western blot showed a single 7.6 kDa band obtained in the induced Rosetta (DE3) pLYsS. ELISA confirmed the molecular identity of the rhIGF-1 epitope, the concentration of purified rhIGF-1 obtained from the ELISA standard curve using rhIGF-1 reference protein as a standard was 300 μg/ml, and activity on WI-38 cells was 2604.17I U/mg.
CONCLUSION
Biologically active native rhIGF-1 protein was successfully expressed. Patents related to the preparation of IGF-1 were mentioned along the text.
Identifiants
pubmed: 35418294
pii: BIOT-EPUB-122487
doi: 10.2174/1872208316666220412105822
doi:
Substances chimiques
IGF1 protein, human
0
Insulin-Like Growth Factor I
67763-96-6
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
266-280Informations de copyright
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