Preparation of a peptide vaccine against GnRH by a bioprocess system based on asparaginase.
GnRH is a promising target in hormone-dependent cancer immunotherapy. In our previous study, we have designed and purified a peptide vaccine GhM (GnRH3-hinge-MVP) by use of the bioprocess system based on asparaginase. Active immunization with GhM in the presence of CFA/IFA evoked strong humoral response. In this study, the motif NRLLLTG with high affinity to nanoparticle carrier VLP HBcDelta-SBD was fused to the C terminus of GhM to form a new peptide vaccine GhMNR (GnRH3-hinge-MVP-NRLLLTG). The fusion protein ansB-C-GhMNR was controlled by vigorous T7lac promotor and expressed effectively as inclusion bodies after induction by lactose and then purified by means of cell disruption, washing and cold ethanol fractionation. After hydrolyzed for 72 h, GhMNR was liberated from the fusion partner ansB-C and purified by CM52 cation exchange chromatography. These results suggested that the bioprocess system is suitable for large-scale expression and purification of the peptide vaccine GhMNR, and even some other proteins or peptides which may be important for industrial or laboratory purposes.
Wang
Xue Jun
XJ
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Nanjing Medical University, Hanzhong Road 140, Nanjing 210029, China. wangxuejun1979728@hotmail.com
Gu
Kai
K
Xu
Jin Shu
JS
Cao
Rong Yue
RY
Li
Ming Hui
MH
Wu
Jie
J
Li
Tai Ming
TM
Liu
Jing Jing
JJ
D020816:Amino Acid Motifs; D000818:Animals; D000917:Antibody Formation; D001215:Asparaginase / Q000737:chemistry*; D019496:Cancer Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D005818:Genetic Engineering; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles; D010957:Plasmids; D011401:Promoter Regions, Genetic; D022223:Vaccines, Subunit / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D019496:Cancer Vaccines; C512006:GnRH3-hinge-MVP; D022223:Vaccines, Subunit; D014764:Viral Proteins; D007987:Gonadotropin-Releasing Hormone; D001215:Asparaginase