Cloning, expression and purification of human epidermal growth factor using different expression systems.
Epidermal growth factor (EGF) is a protein that belongs to the family of growth factors that bind the ErbB receptors, which play a prominent role in the development of carcinomas. We had demonstrated that potato carboxypeptidase inhibitor (PCI) acts as an EGF antagonist. Because of the low affinity of PCI for the epidermal growth factor receptor, it was decided to design EGF mutants with PCI abilities. In order to achieve this we have first cloned, expressed and purified the native protein, EGF. Different expression systems with different locations of the recombinant protein were designed and a purification protocol was designed with those which allowed expression of EGF. Finally, the sample needed folding. Differences in the amount of EGF obtained and its activity were observed depending on the expression system used.
Ferrer Soler
L
L
Unitat de Bioquímica, Departament de Biologia, Facultat de Ciències, Universitat de Girona, Campus de Montilivi, Spain.
Cedano
J
J
Querol
E
E
de Llorens
R
R
D003001:Cloning, Molecular; D017931:DNA Primers; D004815:Epidermal Growth Factor / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor