[Preparation and characterization of monoclonal antibody against Enterohemorrhagic Escherichia coli O157:H7 EspA].
OBJECTIVE
To prepare hybridoma cell lines were obtained by fusing Sp2/0 with spleen cells from BALB/c mice immunized with killed enterohemorrhagic E.coli O157:H7 EspA (EHEC O157:H7 EspA).


METHODS
The subclass isotype and the specificity of monoclonal antibodies (mAb) were determined and identified by ELISA, Western blot and immune fluorescence staining.


RESULTS
Isotype of 3 mAb was IgG1kappa, IgG1lambda, and IgG2kappa, respectively, and the affinity constant were 3.0x10(9), 2.8x10(9), 1.9x10(9). As demonstrated by Western blot, these 3 mAb specifically reacted with EspA protein and EHEC O157:H7. Useing immune fluorescence staining, EHEC O157:H7 could adhere to the membrace of Hela cell.


CONCLUSIONS
Three hybridoma cell lines can stably secrete anti-EspA mAb with high-titer and high-specific have been established. It can be used to deeply study EHEC O157:H7 pathopoiesis mechanism.
Yu
Shu
S
Department of Clinical Microbiology, Third Military Medical University, Chongqing 400038, China.
Luo
Ping
P
Chen
Hong-zhang
HZ
Li
Hai-xia
HX
Mao
Xu-hu
XH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D045744:Cell Line, Tumor; D019453:Escherichia coli O157 / Q000276:immunology*; D005260:Female; D005455:Fluorescent Antibody Technique; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G