[Construction and immunogenicity of hCG chimeric peptides (CP1 and CP10) containing three self- B cell epitopes and six foreign T cell epitopes].
Two bio-synthesizing chimeric peptide (CP) immunogens named CP1 and CP10 have been designed, which consist of three linear B cell epitopes (BCE) of human chorionic gonadotropin beta subunit (beta-hCG) and six foreign T cell epitopes including two "promiscuous" TCEs from hepatitis B surface antigen and tetanus toxoid. Two artificial genes encoding CP1 and its derivative CP10 were synthesized, which could be expressed in E. coli at the level of about 1% of the total cell proteins when inserted into the thermo-induction vector respectively. In Western blot tests, the expressed CP1 and CP10 proteins with about 16.5 kD shown on the SDS-polyacrylamide gel electrophoresis (PAGE) gel can be recognized by the monoclonal or polyclonal antibodies specific to each linear epitope of beta-hCG. Each of expressed proteins can be purified with 95% relative homogeneity using our improved method of preparative gel PAGE. Their yields were about 1-2 mg per 12 L culture. Also, the CPl and CP10 immunogens can induce antibodies in mice that recognize recombinant CP1 betar CP10 and natural beta-hCG, and there are three anti-beta5, beta9 and beta8 BCE antibodies in their antisera. The construction and expression of beta-hCG CP1 and CP10 will provide new immunogens for developing an ideal and superior hCG birth control and/or tumor therapeutic vaccine.
Xu
Wan Xiang
WX
Department of Reproductive Biology, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China. xuwx@sippr.stc.sh.cn
He
Ya Ping
YP
Qiu
De Yi
DY
Liao
Mao Chuan
MC
Hong
Ai Zhen
AZ
Ji
Zhao Neng
ZN
Gu
Shao Hua
SH
Chen
Jin Zhong
JZ
Xie
Yi
Y
D000818:Animals; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000235:genetics / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D051379:Mice; D008957:Models, Genetic; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*
D018997:Chorionic Gonadotropin, beta Subunit, Human; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D011993:Recombinant Fusion Proteins