Granulocyte colony-stimulating factor (GCSF) fused with Fc Domain produced from E. coli is less effective than Polyethylene Glycol-conjugated GCSF

被引：20

作者：

Do, Bich Hang ^{[1
]}

Kang, Hyo Jeong ^{[1
]}

Song, Jung-A ^{[1
]}

Minh Tan Nguyen ^{[1
]}

Park, Sangsu ^{[1
]}

Yoo, Jiwon ^{[1
]}

Anh Ngoc Nguyen ^{[1
]}

Kwon, Grace G. ^{[1
]}

Jang, Jaepyeong ^{[1
]}

Jang, Mihee ^{[1
]}

Lee, Sunju ^{[1
]}

So, Seoungjun ^{[1
]}

Sim, Seongrak ^{[1
]}

Lee, Kyung Jin ^{[2
]}

Osborn, Mark J. ^{[1
,3
]}

Choe, Han ^{[1
]}

机构：

[1] Univ Ulsan, Coll Med, Asan Minnesota Inst Innovating Transplantat, Dept Physiol,Biomed Inst Technol,Asan Med Ctr, Seoul 05505, South Korea

[2] Univ Ulsan, Coll Med, Asan Med Ctr, Dept Convergence Med,Asan Inst Life Sci, Seoul 05505, South Korea

[3] Univ Minnesota, Stem Cell Inst, Div Blood & Marrow Transplantat, Dept Pediat,Ctr Genome Engn, Minneapolis, MN 55455 USA

来源：

SCIENTIFIC REPORTS | 2017年 / 7卷

基金：

新加坡国家研究基金会;

关键词：

G-CSF; ESCHERICHIA-COLI; PEGYLATION; PHARMACOKINETICS; PEGFILGRASTIM;

D O I：

10.1038/s41598-017-06726-7

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Human granulocyte colony-stimulating factor (GCSF) is a well-known cytokine for neutropenia treatment. However, daily injections are required due to the short circulating half-life of the protein. To overcome this bottleneck, we fused GCSF with the Fc domain of IgG1 at the C terminus (GCSF-Fc) and with the maltose binding protein (MBP) tag at the N-terminus and expressed it as a soluble protein in the cytoplasm of E. coli. We also conjugated PEG aldehyde to GCSF to make PEG-GCSF. The bioactivities of GCSF-Fc and PEG-GCSF were similar to native GCSF using the mouse M-NFS-60 myelogenous leukemia cell line. The EC50 dose-response curves for GCSF, GCSF-Fc and PEG-GCSF were 37 +/- 12 pM, 75 +/- 13.5 pM and 46 +/- 5.5 pM, respectively. When the proteins were injected into neutropenic rats, the group injected with PEG-GCSF showed the highest and fastest recovery of neutrophils, followed by GCSF-Fc and GCSF. ELISA assay revealed the PEG-GCSF had the longest plasma circulation (> 72 h), followed by GCSF-Fc (> 48 h) and GCSF (similar to 24 h), which is consistent with the in vivo activities of the proteins. In summary, the GCSF-Fc purified from E. coli was not as efficient as PEG-GCSF in treating neutropenic rats.

引用

页数：9

共 30 条

[1] Pegfilgrastim and daily granulocyte colony-stimulating factor: patterns of use and neutropenia-related outcomes in cancer patients in Spain - results of the LEARN Study [J].

Almenar, D. ;

Mayans, J. ;

Juan, O. ;

Garcia, Bueno J. M. ;

Jalon, Lopez J., I ;

Frau, A. ;

Guinot, M. ;

Cerezuela, P. ;

Garcia, Buscalla E. ;

Gasquet, J. A. ;

Sanchez, J. .

EUROPEAN JOURNAL OF CANCER CARE, 2009, 18 (03) :280-286

[2] Relationship between molecular mass and duration of activity of polyethylene glycol conjugated granulocyte colony-stimulating factor mutein [J].

Bowen, S ;

Tare, N ;

Inoue, T ;

Yamasaki, M ;

Okabe, M ;

Horii, I ;

Eliason, JF .

EXPERIMENTAL HEMATOLOGY, 1999, 27 (03) :425-432

[3] Construction of a set Gateway-based destination vectors for high-throughput cloning and expression screening in Escherichia coli [J].

Busso, D ;

Delagoutte-Busso, B ;

Moras, D .

ANALYTICAL BIOCHEMISTRY, 2005, 343 (02) :313-321

[4] Hematopoietic Properties of Granulocyte Colony-Stimulating Factor/Immunoglobulin (G-CSF/IgG-Fc) Fusion Proteins in Normal and Neutropenic Rodents [J].

Cox, George N. ;

Chlipala, Elizabeth A. ;

Smith, Darin J. ;

Carlson, Sharon J. ;

Bell, Stacie J. ;

Doherty, Daniel H. .

PLOS ONE, 2014, 9 (03)

[5] Enhanced circulating half-life and hematopoietic properties of a human granulocyte colony-stimulating factor/immunoglobulin fusion protein [J].

Cox, GN ;

Smith, DJ ;

Carlson, SJ ;

Bendele, AM ;

Chlipala, EA ;

Doherty, DH .

EXPERIMENTAL HEMATOLOGY, 2004, 32 (05) :441-449

[6]

DEMETRI GD, 1991, BLOOD, V78, P2791

[7] Soluble Prokaryotic Overexpression and Purification of Bioactive Human Granulocyte Colony-Stimulating Factor by Maltose Binding Protein and Protein Disulfide Isomerase [J].

Do, Bich Hang ;

Ryu, Han-Bong ;

Hoang, Phuong ;

Koo, Bon-Kyung ;

Choe, Han .

PLOS ONE, 2014, 9 (03)

[8] Site-specific attachment of functionalized poly(ethylene glycol) to the amino terminus of proteins [J].

Gaertner, HF ;

Offord, RE .

BIOCONJUGATE CHEMISTRY, 1996, 7 (01) :38-44

[9] Transcytosis and catabolism of antibody [J].

Ghetie, V ;

Ward, ES .

IMMUNOLOGIC RESEARCH, 2002, 25 (02) :97-113

[10] Albugranin™, a recombinant human granulocyte colony stimulating factor (G-CSF) genetically fused to recombinant human albumin induces prolonged myelopoietic effects in mice and monkeys [J].

Halpern, W ;

Riccobene, TA ;

Agostini, H ;

Baker, K ;

Stolow, D ;

Gu, ML ;

Hirsch, J ;

Mahoney, A ;

Carrell, J ;

Boyd, E ;

Grzegorzewski, KJ .

PHARMACEUTICAL RESEARCH, 2002, 19 (11) :1720-1729

← 1 2 3 →