Abstract
Objectives
To develop a method for in vitro assembly of recombinant proteins expressed in E. coli into chimeric virus-like particles (cVLPs).
Results
A fusion protein (Bepi-Cap-A) between capsid protein (Cap) of PCV2b and B cell epitope (Bepi) of IBDV was expressed in E. Coli, and purified. For assembling them into cVLPs (Bepi-Cap-VLP), the Bepi-Cap-A was suspended in buffer C [0.03% (“%” stands for “v/v” unless otherwise indicated) polyethylene glycol, 0.4 M Tris, 10 mM β-mercaptoethanol, 5% glycerol, 0.02% (w/v) gellan gum, 0.1 M glycine, 0.03% Tween 80, 500 mM NaCl], and incubated. After centrifugation, the pellet was resuspended in buffer D [50 mM Na2HPO4, 50 mM NaH2PO4, 0.01% (w/v) gellan gum, 0.05 mM EDTA, 500 mM NaCl, 0.03% Tween 80, pH 6.5], and then dialyzed against dialysis buffer (50 mM Na2HPO4, 50 mM NaH2PO4, 500 mM NaCl, 0.03% Tween 80, pH 6.5). The procedure resulted in typical and immunogenic Bepi-Cap-VLP.
Conclusions
The data provide a method which is feasible for in vitro assembly of recombinant proteins into chimeric virus-like particles.
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Acknowledgements
The authors would like to thank Ye Li for technical assistance, Xin Li for checking problems, Peiyin Zhang for technical guidance. We also thank Dr. Yongli Yu to help revise the manuscript.
Supporting information
Supplementary Table 1 The physicochemical difference among Cap and three putative proteins by ProtParam tool.
Supplementary Figure 1 Prediction of B-cell epitopes in the capsid protein VP2 of infectious bursal disease virus (IBDV). (a) Cartoon structures of IBDV VP2. The quaternary structure of VP2 was depicted by SWISS MODEL. The loops of VP2 are magnified 6 x. (b) Location of candidate B-cell epitopes in VP2. Based on their hydrophilicity, flexibility, accessibility and antigenicity, the B-cell epitope in VP2 was predicted in the Protean system, and denoted using a rectangular line. (c) Prediction of linear B-cell epitopes in VP2. The linear B-cell epitopes were predicted by PepiPred 1.0., and denoted with the yellow shadow within the rectangular line.
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In the animal assay, mice were maintained in microisolator cages under laminar flow condition and used for the mouse experiments in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals. All authors are in agreement with the content of the manuscript.
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Sun, X., Xing, S., Wang, S. et al. In vitro assembly of chimeric virus-like particles composed of a porcine circovirus 2b capsid protein and a B-cell epitope of infectious bursal disease virus. Biotechnol Lett 44, 429–438 (2022). https://doi.org/10.1007/s10529-022-03237-y
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DOI: https://doi.org/10.1007/s10529-022-03237-y