Glycosylated and non-glycosylated recombinant human granulocyte colony-stimulating factor (rhG-CSF)—what is the difference?
Two forms of recombinant human G-CSF (rhG-CSF) are available for clinical use: filgrastim is expressed inE coli and non-glycosylated, whereas lenograstim is derived from Chinese hamster ovary (CHO) cells and glycosylated. The function of the sugar chain, accounting for approximately 4% of the molecular weight of lenograstim (and native G-CSF), is not known. Glycosylation of the G-CSF molecule does not prolong its circulation half life. Lenograstim is more active than filgrastim (and research-use deglycosylated G-CSF) on a weight-by-weight basis inin vitro colony-forming and cell line assays. An international potency standard assigns a specific activity of 100 000 IU/μg to filgrastim and 127 760 IU/μg to lenograstim. Correspondingly, two randomised crossover studies in normal subjects, comparingmass equivalent doses of the two rhG-CSFs, have demonstrated a 25–30% higher concentration of blood stem cells (CD34+, CFU-GM) during lenograstim administration. No difference in side effects was observed. Results from a prospective, randomised, non-crossover trial in breast cancer patients suggest thatbioequivalent doses of filgrastim and lenograstim have a similar effect on mobilisation of CD34+ cells and immature CD34+ cell subsets, respectively. Although comparisons outside the setting of stem cell mobilisation are lacking, the clinical relevance of the greater specific activity of lenograstim may thus be limited. The difference in potency between μg identical doses of the two rhG-CSFs makes dosing in biological units (IU) rather than mass units (μg) more appropriate.
KeywordsG-CSF glycosylation filgrastim lenograstim comparative study
Asano S. Human granulocyte colony-stimulating factor: its basic aspects and clinical applications.Am J Pediatr Hematol Oncol
: 400–413.PubMedCrossRefGoogle Scholar
Demetri GD, Griffin JD. Granulocyte colony-stimulating factor and its receptor.Blood
: 2791–2808.PubMedGoogle Scholar
Souza Let al.
Recombinant human granulocyte colony-stimulating factor: Effects on normal and leukemic myeloid cells.Science
: 61.PubMedCrossRefGoogle Scholar
Nagata Set al.
Molecular cloning and expression of cDNA for human granulocyte colony-stimulating factor.Nature
: 415–418.PubMedCrossRefGoogle Scholar
Welte K, Gabrilove J, Bronchud MH, Platzer E, Morstyn G. Filgrastim (r-metHuG-CSF): the first 10 years.Blood
: 1907–1929.PubMedGoogle Scholar
To LB, Haylock DN, Simmons PJ, Juttner CA. The biology and clinical uses of blood stem cells.Blood
: 2233–2258.PubMedGoogle Scholar
Höglund Met al.
Mobilisation of CD34+
cells by glycosylated and nonglycosylated G-CSF in healthy volunteers—a comparative study.Eur J Haematol
: 177–183.PubMedCrossRefGoogle Scholar
Watts MJ, Addison I, Long SG, Hartley S, Warrington S, Boyce M, Linch DC. Crossover study of the haematological effects and pharmacokinetics of glycosylated and non-glycosylated G-CSF in healthy volunteers.Br J Haematol
: 474–479.PubMedCrossRefGoogle Scholar
de Arriba Fet al.
Prospective randomized study comparing the efficacy of bioequivalent doses of glycosylated and nonglycosylated rG-CSF for mobilizing peripheral blood progenitor cells.Br J Haematol
: 418–420.PubMedCrossRefGoogle Scholar
Linch DCet al.
Randomised vehicle-controlled dose finding study of glycosylated recombinant human granulocyte colony-stimulating factor after bone marrow transplantation.Bone Marrow Transplant
: 307–311.PubMedGoogle Scholar
Weaver CHet al.
Mobilization and harvesting of peripheral blood stem cells: randomised evaluations of different doses of filgrastim.Br J Haematol
: 338–347.PubMedCrossRefGoogle Scholar
Kroger Net al.
10 μg Versus 2×5μg Versus 2×5 μg G-CSF in steady state mobilization of CD34+
progenitor cells in high risk breast cancer patients: higher yield by splitting the dose.Blood
(suppl 1): 592a.Google Scholar
Metcalf D. The colony stimulating factors.Cancer
: 2185–2195.PubMedCrossRefGoogle Scholar
Kolvenbach CG, Langley KE, Strickland TW, Kenney WC, Arakawa T. Densimetric determination of carbohydrate content in glycoproteins.J Biochem Biophys Meth
: 295–300.PubMedCrossRefGoogle Scholar
Wang C, Eufemi M, Turano C, Giartosio A. Influence of the carbohydrate moiety on the stability of glycoproteins.Biochemistry
: 7299–7307.PubMedCrossRefGoogle Scholar
Fukuda MN, Sasaki H, Lopez L, Fukuda M. Survival of recombinant erythropoietin in the circulation: the role of carbohydrates.Blood
: 84–89.PubMedGoogle Scholar
Hovgaard D, Mortensen BT, Schifter S, Nissen NI. Comparative pharmacokinetics of single-dose administration of mammalian and bacterially-derived recombinant human granulocyte-macrophage colony-stimulating factor.Eur J Haematol
: 32–36.PubMedCrossRefGoogle Scholar
Cebon Jet al.
Granulocyte-macrophage colony stimulating factor from human lymphocytes. The effect of glycosylation on receptor binding and biological activity.J Biol Chem
: 4483–4491.PubMedGoogle Scholar
Gribben JGet al.
Development of antibodies to unprotected glycosylation sites on recombinant human GM-CSF [see comments].Lancet
: 434–437.PubMedCrossRefGoogle Scholar
Ragnhammar Pet al.
Induction of anti-recombinant human granulocyte-macrophage colony-stimulating factor (Escherichia coli
-derived) antibodies and clinical effects in nonimmuno-compromised patients.Blood
: 4078–4087.PubMedGoogle Scholar
Moonen P, Mermod JJ, Ernst JF, Hirschi M, DeLamarter JF. Increased biological activity of deglycosylated recombinant human granulocyte/macrophage colony-stimulating factor produced by yeast or animal cells.Proc Natl Acad Sci USA
: 4428–4431.PubMedCrossRefGoogle Scholar
Kubota Net al.
Structural characterization of natural and recombinant human granulocyte colony-stimulating factors.J Biochem (Tokyo)
: 486–492.Google Scholar
Arakawa T, Prestrelski SJ, Narhi LO, Boone TC, Kenney WC. Cysteine 17 of recombinant human granulocytecolony stimulating factor is partially solvent-exposed.J Protein Chem
: 525–531.PubMedCrossRefGoogle Scholar
Hill CP, Osslund TD, Eisenberg D. The structure of granulocyte-colony-stimulating factor and its relationship to other growth factors.Proc Natl Acad Sci USA
: 5167–5171.PubMedCrossRefGoogle Scholar
Oh-eda M, Hase S, Ono M, Ikenaka T. Structures of the sugar chains of recombinant human granulocyte-colony-stimulating factor produced by Chinese hamster ovary cells.J Biochem (Tokyo)
: 544–546.Google Scholar
Christi L, Clogston CL, Hu S, Boone TC, Lu HS. Glycosidase digestion, electrophoresis and chromatographic analysis of recombinant human granulocyte colony-stimulating factor glycoforms produced in Chinese hamster ovary cells.J Chromatogr
: 55–62.CrossRefGoogle Scholar
Oh-eda Met al.
Preparation of pyridylaminated O-linked sugar chains from glycoproteins blotted on a polyvinylidene difluoride membrane and application to human granulocyte colony-stimulating factor.Analytical Biochemistry
: 369–371.PubMedCrossRefGoogle Scholar
Nissen C, Dalle-Carbonare Y, Moser Y.In vitro
comparison of the biological potency of glycosylated versus non-glycosylated rG-CSF.Drug Invest
: 346–352.Google Scholar
Stute Net al.
Pharmacokinetics of subcutaneous recombinant human granulocyte colony-stimulating factor in children.Blood
: 2849–2854.PubMedGoogle Scholar
Pedrazzoli Pet al.
Effects of glycosylated and non-glycosylated G-CSFs, alone and in combination with other cytokines, on the growth of human progenitor cells.Anticancer Res
: 1781–1785.PubMedGoogle Scholar
Decleva E, Cramer R, Zabucchi G. Glycosylation improves the priming effect exerted by recombinant human granulocyte colony-stimulating factor (lenograstim) on human neutrophil superoxide production.Int J Tissue React
: 191–198.PubMedGoogle Scholar
Mire-Sluis AR, Das RG, Thorpe R. The international standard for granulocyte colony stimulating factor (G-CSF). Evaluation in an international collaborative study.J Immunol Methods
: 117–126.PubMedCrossRefGoogle Scholar
Okabe Met al. In vitro
hematopoietic effect of mutant human granulocyte colony-stimulating factor.Blood
: 1788–1793.PubMedGoogle Scholar
Gisselbrecht Cet al.
A phase III randomised placebo-controlled study of lenograstim (glycosylated rHu-G-CSF) in 315 paediatric and adult autologous or allogeneic bone marrow transplant patients.Lancet
: 696.PubMedCrossRefGoogle Scholar
Tanaka Het al.
Three types of recombinant human granulocyte colony-stimulating factor have equivalent biological activities in monkeys.Cytokine
: 360–369.PubMedCrossRefGoogle Scholar
Nohynek GJ, Plard JP, Wells MY, Zerial A, Roquet F. Comparison of the potency of glycosylated and non-glycosylated recombinant human granulocyte colony-stimulating factors in neutropenic and non-neutropenic CD rats.Cancer Chemother Pharmacol
: 259–266.PubMedCrossRefGoogle Scholar
Roberts AWet al.
Broad inter-individual variations in circulating progenitor cell numbers induced by granulocyte colony-stimulating factor therapy.Stem Cells
: 512–516.PubMedCrossRefGoogle Scholar
Stroncek Det al.
Treatment of normal individual with granulocyte-colony-stimulating factor: Donor experiences and the effects on peripheral blood CD34+ cell counts on the collection of peripheral blood stem cells.Transfusion
: 601.PubMedCrossRefGoogle Scholar
Denzlinger Cet al.
Differential activation of the endogenous leukotriene biosynthesis by two different preparations of granulocyte-macrophage colony-stimulating factor in healthy volunteers.Blood
: 2007–2013.PubMedGoogle Scholar
Bonilla MAet al.
Long-term safety of treatment with recombinant human granulocyte colony-stimulating factor (r-metHuG-CSF) in patients with severe congenital neutropenias.Br J Haematol
: 723–730.PubMedGoogle Scholar
Donadieu Jet al.
A European phase II study of recombinant human granulocyte colony-stimulating factor (lenograstim) in the treatment of severe chronic neutropenia in children.Eur J Pediatr
: 693–700.PubMedCrossRefGoogle Scholar
Oh-Eda Met al.
O-linked sugar chain of human granulocyte colony-stimulating factor protects against polymerization and denaturation allowing it to retain its biological activity.J Biol Chem
: 11432–11435.PubMedGoogle Scholar
Kishita Met al.
Stability of G-CSF in serum.Clin Rep
: 1–4.Google Scholar
Mire-Sluis AR. Activity on cell lines: Stability of rG-CSFs in human serum.Int J Hematol
: S11-S12.Google Scholar
Gervais V, Zerial A, Oschkinat H. NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor.Eur J Biochem
: 386–395.PubMedCrossRefGoogle Scholar
Höglund M, Smedmyr B, Simonsson B, Tötterman T, Bengtsson M. Dose-dependent mobilisation of haematopoietic progenitor cells in healthy volunteers receiving glycosylated rHuG-CSF.Bone Marrow Transplant
: 19–27.PubMedGoogle Scholar
Kulkarni Set al.
Comparison of equal doses of lenograstim and filgrastim for mobilization of blood stem cells in patients with hematologic malignancies.Blood
(suppl 1): 4226a.Google Scholar
Saccardi R, Avanci G, Bezzini R. Mobilization of PBSC for hematological rescue: comparison between glycosylated and non-glycosylated G-CSF.Bone Marrow Transplant
(Suppl. 1): S11.Google Scholar