[Synthesis of dominant epitopes on N-terminal part of BPI and preparation of corresponding antisera].
OBJECTIVE
To synthesize B cell dominant epitopes on N-terminal part of bactericidal/permeability increasing protein (BPI) and prepare the corresponding antisera.


METHODS
The antigenicity, hydrophilicity, flexibility, surface probability and secondary structure of N-terminal amino acids 1-199 on BPI were predicted by bioinformatics applications. Two antigen peptides TA/IK were designed and synthesized on the basis of the above analysis. Then the TA/IK were respectively conjugated to keyhole limpet hemocyanin (KLH) and injected into rabbits to prepare corresponding antisera. Indirect ELISA was performed to analyze the antigenicity of TA/IK and to test the titer of the antisera. And Western blot was used to identify the specificity of the antisera.


RESULTS
(1) Two B cell epitope-based peptides TA/IK were successfully synthesized; (2) the peptides could bind to commercial polyclonal antibody, anti-BPI(55); (3) titers of the antisera against TA/IK were up to 1:51,200, 1:25,600, respectively; (4) Western blot analysis revealed that these antisera could specifically react with the standard sample of BPI(55).


CONCLUSIONS
The two synthetic antigen peptides TA/IK are indeed dominant epitopes of BPI N-terminal part, and the corresponding antisera are competent for detecting and identifying the N-terminal fragments of BPI.
Chen
Yan-qiu
YQ
Department of Immunology, Capital Medical University, Beijing 100069, China.
Peng
Zhi
Z
Long
Jun
J
Zhang
Guo-min
GM
Lu
Zhe
Z
An
Yun-qing
YQ
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D001798:Blood Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D015153:Blotting, Western; D019295:Computational Biology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D007106:Immune Sera / Q000096:biosynthesis*; D008297:Male; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D000939:Epitopes; D007106:Immune Sera; D010455:Peptides; C021623:bactericidal permeability increasing protein