Generation of streptavidin-tagged human-granulocyte macrophage colony-stimulating factor fusion proteins / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1389-1393, 2012.
Article
Dans Anglais
| WPRIM
| ID: wpr-315456
ABSTRACT
<p><b>OBJECTIVE</b>To obtain streptavidin-tagged human granulocyte-macrophage colony-stimulating factor (SA/hGM-CSF) fusion protein and evaluate its bioactivity .</p><p><b>METHODS</b>PET24a-6His-SA-L-hGM-CSF and PET24a-hGM-CSF-L-SA-6His plasmids were constructed and expressed in Rosetta (DE3) host bacteria to generate the fusion proteins. The two fusion proteins were refolded by gradient dialysis after Ni-NTA affinity chromatography and finally purified using DEAE-sepharose FF anion exchange chromatography. MTT method was used to evaluate the effect of SA/hGM-CSF fusion proteins in inducing the proliferation of human erythroleukemia cells (TF-1). The efficiency of the fusion proteins for surface modification of biotinylated MB49 tumor cells was evaluated by flow cytometry.</p><p><b>RESULTS</b>The recombinant fusion proteins SA-hGM-CSF and hGM-CSF-SA were highly expressed in Rosetta (DE3) at about 20% of the total bacterial proteins, with a purity of about 96% after purification. The two fusion proteins exhibited bifunctional activities, namely the pro-proliferation effect on human erythroleukemia cells (TF-1) and SA-mediated high-affinity binding to biotinylated cell surfaces (with an anchoring modified rate of about 99%).</p><p><b>CONCLUSION</b>SA/hGM-CSF bi-fusion proteins obtained in this study lays the groundwork for the development of cancer cell vaccines with surface modification by hGM-CSF.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Pharmacologie
/
Plasmides
/
Marqueurs biologiques
/
Chimie
/
Facteur de stimulation des colonies de granulocytes et de macrophages
/
Vaccins anticancéreux
/
Streptavidine
/
Lignée cellulaire tumorale
/
Diterpènes
/
Escherichia coli
Limites du sujet:
Humains
langue:
Anglais
Texte intégral:
Journal of Southern Medical University
Année:
2012
Type:
Article
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