Human granulocyte-colony stimulating factor (G-CSF)/stem cell factor (SCF) fusion proteins: design, characterization and activity.
Gitana MickieneIndre DalgedieneGintautas ZvirblisZilvinas DapkunasIeva PlikusieneErnesta Buzavaite-VertelieneZigmas BalevičiusAudronė RukšėnaitėMilda PleckaitytePublished in: PeerJ (2020)
The novel SCF-Lα-GCSF and GCSF-Lα-SCF proteins were synthesized in Escherichia coli. The purity of the heterodimers reached >90% as determined by RP-HPLC. The identity of the proteins was confirmed using the Western blot and HPLC/ESI-MS assays. An array of multimeric forms, non-covalently associated dimers or trimers were detected in the protein preparations by SE-HPLC. Each protein induced a dose-dependent proliferative response on the cell lines. At equimolar concentration, the heterodimers retain 70-140% of the SCF monomer activity (p ≤ 0.01) in promoting the M-07e cells proliferation. The G-CSF moiety in GCSF-Lα-SCF retained 15% (p ≤ 0.0001) and in SCF-Lα-GCSF retained 34% (p ≤ 0.01) of the monomeric G-CSF activity in stimulating the growth of G-NFS-60 cells. The obtained results were in good agreement with the binding data of each moiety in the fusion proteins to their respective receptors. The increase in the absolute neutrophil count in rats caused by the SCF-Lα-GCSF protein corresponded to the increase induced by a mixture of SCF and G-CSF.
Keyphrases
- ms ms
- induced apoptosis
- escherichia coli
- stem cells
- simultaneous determination
- mass spectrometry
- high throughput
- cell cycle arrest
- binding protein
- multiple sclerosis
- cell proliferation
- peripheral blood
- solid phase extraction
- electronic health record
- oxidative stress
- staphylococcus aureus
- machine learning
- tandem mass spectrometry
- high resolution
- cerebrospinal fluid
- multidrug resistant
- drug induced
- small molecule
- mesenchymal stem cells
- high density
- molecularly imprinted
- biofilm formation
- stress induced
- diabetic rats
- pluripotent stem cells