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

Bich Hang Do; Hyo Jeong Kang; Jung A. Song; Minh Tan Nguyen; Sangsu Park; Jiwon Yoo; Anh Ngoc Nguyen; Grace G. Kwon; Jaepyeong Jang; Mihee Jang; Sunju Lee; Seoungjun So; Seongrak Sim; Kyung Jin Lee; Mark J. Osborn; Han Choe

doi:10.1038/s41598-017-06726-7

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

Bich Hang Do, Hyo Jeong Kang, Jung A. Song, Minh Tan Nguyen, Sangsu Park, Jiwon Yoo, Anh Ngoc Nguyen, Grace G. Kwon, Jaepyeong Jang, Mihee Jang, Sunju Lee, Seoungjun So, Seongrak Sim, Kyung Jin Lee, Mark J. Osborn, Han Choe

Pediatrics - Blood/Marrow Transplant

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

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 (∼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.

Original language	English (US)
Article number	6480
Journal	Scientific reports
Volume	7
Issue number	1
DOIs	https://doi.org/10.1038/s41598-017-06726-7
State	Published - Dec 1 2017

Bibliographical note

Funding Information:
We thank Dr. Dok Hyun Yoon for his invaluable advice on the interpretation of the in vivo experiment result. This study was supported by grants (NRF-2015K1A4A3046807, 2008-0062286, and 2017M3A9F8031039) from the National Research Foundation of Korea and a grant (2017-307) from the Asan Institute for Life Sciences, Seoul, Korea.

Publisher Copyright:
© 2017 The Author(s).

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access

10.1038/s41598-017-06726-7

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5526978

OpenUrl availability

Full text

Cite this

Do, B. H., Kang, H. J., Song, J. A., Nguyen, M. T., Park, S., Yoo, J., Nguyen, A. N., Kwon, G. G., Jang, J., Jang, M., Lee, S., So, S., Sim, S., Lee, K. J., Osborn, M. J., & Choe, H. (2017). Granulocyte colony-stimulating factor (GCSF) fused with Fc Domain produced from E. coli is less effective than Polyethylene Glycol-conjugated GCSF. Scientific reports, 7(1), Article 6480. https://doi.org/10.1038/s41598-017-06726-7

Do, BH, Kang, HJ, Song, JA, Nguyen, MT, Park, S, Yoo, J, Nguyen, AN, Kwon, GG, Jang, J, Jang, M, Lee, S, So, S, Sim, S, Lee, KJ, Osborn, MJ & Choe, H 2017, 'Granulocyte colony-stimulating factor (GCSF) fused with Fc Domain produced from E. coli is less effective than Polyethylene Glycol-conjugated GCSF', Scientific reports, vol. 7, no. 1, 6480. https://doi.org/10.1038/s41598-017-06726-7

@article{710c357a37734a08923fa3809ba5b631,

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

abstract = "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 (∼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.",

author = "Do, {Bich Hang} and Kang, {Hyo Jeong} and Song, {Jung A.} and Nguyen, {Minh Tan} and Sangsu Park and Jiwon Yoo and Nguyen, {Anh Ngoc} and Kwon, {Grace G.} and Jaepyeong Jang and Mihee Jang and Sunju Lee and Seoungjun So and Seongrak Sim and Lee, {Kyung Jin} and Osborn, {Mark J.} and Han Choe",

note = "Funding Information: We thank Dr. Dok Hyun Yoon for his invaluable advice on the interpretation of the in vivo experiment result. This study was supported by grants (NRF-2015K1A4A3046807, 2008-0062286, and 2017M3A9F8031039) from the National Research Foundation of Korea and a grant (2017-307) from the Asan Institute for Life Sciences, Seoul, Korea. Publisher Copyright: {\textcopyright} 2017 The Author(s).",

year = "2017",

month = dec,

day = "1",

doi = "10.1038/s41598-017-06726-7",

language = "English (US)",

volume = "7",

journal = "Scientific reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Granulocyte colony-stimulating factor (GCSF) fused with Fc Domain produced from E. coli is less effective than Polyethylene Glycol-conjugated GCSF

AU - Do, Bich Hang

AU - Kang, Hyo Jeong

AU - Song, Jung A.

AU - Nguyen, Minh Tan

AU - Park, Sangsu

AU - Yoo, Jiwon

AU - Nguyen, Anh Ngoc

AU - Kwon, Grace G.

AU - Jang, Jaepyeong

AU - Jang, Mihee

AU - Lee, Sunju

AU - So, Seoungjun

AU - Sim, Seongrak

AU - Lee, Kyung Jin

AU - Osborn, Mark J.

AU - Choe, Han

N1 - Funding Information: We thank Dr. Dok Hyun Yoon for his invaluable advice on the interpretation of the in vivo experiment result. This study was supported by grants (NRF-2015K1A4A3046807, 2008-0062286, and 2017M3A9F8031039) from the National Research Foundation of Korea and a grant (2017-307) from the Asan Institute for Life Sciences, Seoul, Korea. Publisher Copyright: © 2017 The Author(s).

PY - 2017/12/1

Y1 - 2017/12/1

N2 - 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 (∼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.

AB - 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 (∼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.

UR - http://www.scopus.com/inward/record.url?scp=85026255858&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85026255858&partnerID=8YFLogxK

U2 - 10.1038/s41598-017-06726-7

DO - 10.1038/s41598-017-06726-7

M3 - Article

C2 - 28744022

AN - SCOPUS:85026255858

SN - 2045-2322

VL - 7

JO - Scientific reports

JF - Scientific reports

IS - 1

M1 - 6480

ER -

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

Abstract

Bibliographical note

UN SDGs

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this