Asparaginase display of human cholesteryl ester transfer protein (CETP) B cell epitopes for inducing high titers of anti-CETP antibodies in vivo.
The recombinant chimeric enzyme, AnsB-TTP-CETPC, comprising asparaginase, tetanus toxin helper T cell epitope and human CETP B cell epitope was expressed as a soluble protein in Escherichia coli. The purified chimeric enzyme exhibited approximate 83% activity of the native asparaginase. After immunization with three doses of chimeric enzyme, high titers of anti-CETP antibodies were induced and lasted more than eighteen weeks in mice, and could even be detected at a dilution of 1:12800 by normal ELISA assay. The specificity of anti-CETP antibody was verified by Western blot assay. After displaying on the surface of asparaginase, the weak antigenicity of CETP epitope was effectively overcome, there after a strong CETP-specific immune response was evoked in mice immunized with the chimeric enzyme. Histochemical analysis of mice kidney tissue showed that immunization with the chimeric enzyme did not cause any pathological changes in mice. Collectively, the chimeric enzyme may be further developed as a vaccine against atherosclerosis in the future.
Gaofu
Qi
Q
Biopharmaceutical College, China Pharmaceutical University, Tongjiaxiang 24, Nanjing 210009, Jiangsu, P.R. China.
Rongyue
Cao
C
Dan
Mao
M
Xiuyun
Zhao
Z
Xuejun
Wang
W
Jie
Wu
W
Jingjing
Liu
L
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000276:immunology*; D001215:Asparaginase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D053480:Cholesterol Ester Transfer Proteins; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D017186:Titrimetry
D000906:Antibodies; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D018985:Epitopes, B-Lymphocyte; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins; D001215:Asparaginase