Large‐scale production, purification, and function of a tumor multi‐epitope vaccine: Peptibody with bFGF/VEGFA
Authors: Xi Chen
Chengcheng Jiang
Ligang Zhang
Hui Zhong
Qingfeng Dong
Sheng Xiong
Ning Deng
Jiangtao Gu
Qifang Song
Jin-Chen Yu
Journal: Engineering in Life Sciences (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Peptibody; bFGF/VEGFA; fermentation; purification; tumor angiogenesis.
Affiliations: Guangdong Province Engineering Research Center for Antibody Drug and Immunoassay Department of Biology Jinan University  Guangzhou P. R. China, The Biomedicine Translational Institute in Jinan University  Guangzhou P. R. China, Guangdong Jida Genetic Medicine Engineering Research Center Co. Ltd  Guangzhou P. R. China, Bio‐Thera Solution Co. Ltd  Guangzhou P. R. China

Abstract:  In tumor tissue, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor A (VEGFA) promote tumorigenesis by activating angiogenesis, but targeting single factor may produce drug resistance and compensatory angiogenesis. The Peptibody with bFGF/VEGFA was designed to simultaneously blockade these two factors. We were aiming to produce this Fc fusion protein in a large scale. The biological characterizations of Peptibody strains were identified as Escherichia coli and the fermentation mode was optimized in the shake flasks and 10-L bioreactor. The fermentation was scaled up to 100 L, with wet cell weight (WCW) 126 g/L, production 1.41 g/L, and productivity 0.35 g/(L·h) of IPTG induction. The target protein was isolated by cation-exchange, hydrophobic and Protein A chromatography, with total recovery of 60.28% and HPLC purity of 86.71%. The host cells protein, DNA, and endotoxin residues were within the threshold. In mouse model, immunization of Peptibody vaccine could significantly suppressed the tumor growth and angiogenesis, with inhibition rate of 57.73 and 39.34%. The Peptibody vaccine could elicit high-titer anti-bFGF and anti-VEGFA antibodies, which inhibited the proliferation and migration of Lewis lung cancer cell cells by decreasing the Akt/MAPK signal pathways. Therefore, the Peptibody with bFGF/VEGFA might be used as a therapeutic tumor vaccine. The large-scale process we developed could support its industrial production and pre-clinical study in the future. 

Retracted: No

PubPeer comments:
 March 2016 Correction for errors in Figures 3 and 5, found here: https: journals.plos.org plospathogens article?id=10.1371 journal.ppat.1005504  *The authors would like to correct Figs 3 and 5, as errors were introduced in the preparation of these figures for publication. In Fig 3B, phosphorylation of JNK was cropped from a large experiment as shown in the uncropped blot in S1 Fig. The cropping for p-JNK and actin were performed incorrectly, shifted over to the right by one lane. The correct p-JNK and actin blots have been inserted into the corrected Fig 3. The raw blots for Fig 3B are presented below in S1 Fig. In Fig 5C, the authors determined that the wrong panel of the actin blot was inserted. To correct this error, the authors performed a new experiment exactly as performed in the original manuscript and present the results in the corrected Fig 5. The raw blots for Fig 5C are presented below in S2 Fig. The corrected version of Figs 3 and 5 can be viewed here.* Here are the original Figures 3B and 5C, compared. The  errors introduced in the figures involved not just using the same blot twice (lanes marked with red arrows), but mirroring it, resizing it, changing the contrast, cutting out one lane and adding it to a different position (blue arrows), and cutting out another lane and adding it to the other end of the blot (green arrows). Could the authors please clarify how these errors occurred? ![file] ,