Co-expression Construction of Qβ Phage Virus-like Particles Presenting Human Interleukin-13 Antigenic Peptide

doi:10.13523/j.cb.20180509

China Biotechnology

2018, Vol. 38

Issue (5): 66-72 DOI: 10.13523/j.cb.20180509

Co-expression Construction of Qβ Phage Virus-like Particles Presenting Human Interleukin-13 Antigenic Peptide

Hong-mei BAI,Wei-wei HUANG,Cun-bao LIU,Wen-jia SUN,Xu YANG,Yan-bing MA(

)

Institute of Medical Biology,Chinese Academy of Medical Science & Peking Union Medical College,Kunming 650118,China

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Abstract

Objective:To construct Qβ phage virus-like particles (VLPs) vaccines presenting human interleukin-13 (IL-13) antigen peptide.Methods:The human IL-13 antigen peptide was genetically recombined into the C terminus of the Qβ phage capsid protein (CP). In BL21 bacteria, the native CP and the recombinant CP with C terminal fused with IL-13 antigen peptide (CP / IL-13) were induced simultaneously by IPTG. VLPs were purified by ammonium sulfate precipitation and sucrose density gradient centrifugation, and the presence of chimeric VLPs was analyzed. The purity of VLPs was analyzed by HPLC and the morphology of the particles was observed by electron microscopy. The mice were subcutaneously immunized with VLPs and sera were collected for detection of human IL-13-specific IgG antibodies by ELISA. RESULTS: The recombinant protein CP and CP / IL-13 were successfully expressed. Both of them appeared in the same fractions of collected samples from density gradient centrifugation and had the same sedimentation behavior as the native Qβ VLPs, while CP / IL-13 alone had no Qβ particle behavior. After purification, high purity particles were obtained. The chimeric particles were similar in shape to Qβ particles. In addition, the VLPs vaccine induced mice to develop an IL-13-specific antibody response.Conclusion:The co-expression strategy can successfully construct chimeric VLPs presenting human IL-13 epitopes, and provide a vaccine form with potentials for clinical application antagonizing the pathological effects of IL-13 in severe human diseases.

Key words： Interleukin-13 Virus-like particles Vaccine

Received: 14 January 2018 Published: 05 June 2018

ZTFLH:

Q819

Corresponding Authors: Yan-bing MA E-mail: may@imbcams.com.cn

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	Hong-mei BAI
	Wei-wei HUANG
	Cun-bao LIU
	Wen-jia SUN
	Xu YANG
	Yan-bing MA

Cite this article:

Hong-mei BAI,Wei-wei HUANG,Cun-bao LIU,Wen-jia SUN,Xu YANG,Yan-bing MA. Co-expression Construction of Qβ Phage Virus-like Particles Presenting Human Interleukin-13 Antigenic Peptide. China Biotechnology, 2018, 38(5): 66-72.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180509 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I5/66

Table 1 Human IL-13 antigen peptide nucleotide sequences and PCR amplification primers

Fig.1 pRSFDuet1 plasmid map

Fig.2 The expression and density gradient centrifugation analysis of the recombinant proteins (a),(c)The co-expression of CP and CP / IL-13 M:Protein marker;1:pRSFDuet1-CP- CP/IL-13 or pRSFDuet1- CP/IL-13-CP without the induction of IPTG;2: pRSFDuet1-CP- CP/IL-13 or pRSFDuet1- CP/IL-13-CP induced by IPTG for 3h;3:The supernatant of induced bacterial cells;4:The precipitate of induced bacterial cells (a)pRSFDuet1-CP- CP/IL-13 (c) pRSFDuet1- CP/IL-13-CP (b),(d) Sucrose density gradient centrifugation analysis of recombinant CP- CP/IL-13 or CP/IL-13-CP M:Protein marker;1-12:The sample fractions from the sucrose density gradient centrifugation collected from top to bottom (b)pRSFDuet1-CP- CP/IL13 (c)pRSFDuet1- CP/IL13-CP (e) Sucrose density gradient centrifugation analysis of CP M:Protein marker;1-12:The sample fractions from the sucrose density gradient centrifugation collected from top to bottom

Fig.3 HPLC analysis of recombinant protein and electronic microscopy of VLPs (a) HPLC profile of recombinant CP/IL-13-CP (b) HPLC profile of recombinant CP (c) Electronic microscopy of CP/IL-13- CPVLPs

Fig.4 Determination of IL-13 antibody levels in sera of the immunized mice by ELISA


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