Transient expression of Fc-fused human glycoprotein 130 in Expi293F suspension cells.
Human glycoprotein 130 (gp130) is a signal-transducing receptor for interleukin 6 (IL-6), whose signaling plays a critical role in chronic inflammation and cancer. The soluble form of gp130 specifically inhibits IL-6 trans-signaling. However, achieving high-level expression of a large glycoprotein such as gp130 is difficult. Here, we designed and constructed one Fc-gp130-pcDNA mammalian expression vector, with the mouse IgG2a Fc fragment added to the N-terminus of human gp130, which greatly increased the secretion of recombinant gp130 protein from Expi293F suspension cells. Recombinant fusion Fc-gp130 was easily and efficiently purified from the supernatant of transfected cells by one-step affinity chromatography. Moreover, Fc-gp130 could automatically form dimers by the disulfide bond. Fc-gp130 was confirmed as a more efficient IL-6 trans-signaling blocker by its higher biological activity against signal transducer and activator of transcription 3 (STAT3). This purified active Fc-gp130 could be used to develop valuable therapeutic agents against inflammatory diseases and cancers.
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China.
Xiaozhi
X
Zhao
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China.
Wei
W
Chen
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China.
Liyuan
L
Ge
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China.
Wei
W
Jiang
Vazyme Biotech Co., Ltd, Nanjing 210000, Jiangsu, PR China.
Bo
B
Tang
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China.
Qing
Q
Zhang
Vazyme Biotech Co., Ltd, Nanjing 210000, Jiangsu, PR China.
Xiaoyu
X
Xu
Vazyme Biotech Co., Ltd, Nanjing 210000, Jiangsu, PR China.
Chong
C
Wang
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, Jiangsu, PR China; Vazyme Biotech Co., Ltd, Nanjing 210000, Jiangsu, PR China. Electronic address: caolin@njau.edu.cn.
Lin
L
Cao
Department of Urology, Drum Tower Hospital, Medical School of Nanjing University, Institute of Urology, Nanjing University, Nanjing 210008, Jiangsu, PR China. Electronic address: dr.guohongqian@gmail.com.
Hongqian
H
Guo
D002460:Cell Line; D050822:Cytokine Receptor gp130 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D050822:Cytokine Receptor gp130
Expi293F; Fc fragment; Glycoprotein 130; Transient expression