title: Efficient Purification of rhG-CSF and its PEGylated Forms and Evaluation for In Vitro Activities.
abstract: Granulocyte-colony stimulating factor (G-CSF) has commonly been used to help the patients to recover from neutropenia inflicted due to radiotherapy, organ transplants and chemotherapy. As the number of people undergoing these therapies and procedures are increasing world-wide, the need for more economical ways of G-CSF production and improvement in its efficacy has become increasingly crucial. In the present study, recombinant human G-CSF (rhG-CSF) was expressed in E. coli and its purification process was optimized by demonstrating better efficiency and higher recoveries (upto 54%) in a multi-step chromatographic purification process, which is greater than the existing reports. Additionally, the efficacy of rhG-CSF was increased by derivatizing with polyethylene glycol (PEG; upto 85% PEGylation), which increases the plasma clearance time, reduces the immunogenicity and requires less frequent administration to the patient. Overall, the present study suggests a cost-effective purification process of rhG-CSF and also proposes its efficient conjugation with PEG for enhanced efficacy as compared to the existing commercially available forms.
authors.forename: Dileep
authors.initials: D
authors.lastname: Tiwari
authors.forename: Shafiul
authors.initials: S
authors.lastname: Haque
authors.forename: Arshad
authors.initials: A
authors.lastname: Jawed
authors.forename: Maheshwari K
authors.initials: MK
authors.lastname: Mishra
authors.forename: Thavendran
authors.initials: T
authors.lastname: Govender
authors.affiliation: Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa. Tiwari@ukzn.ac.za.
authors.forename: Hendrik G
authors.initials: HG
authors.lastname: Kruger
mesh_terms: D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D011092:Polyethylene Glycols / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D011092:Polyethylene Glycols; C000595209:polyethylene glycol 1000