[Preparation of specific antibodies against recombinant Eml8 antigen and detection of the circulating antigen].
OBJECTIVE
To prepare specific antibodies against the recombinant Eml8 antigen (ReEml8), establish a sandwich ELISA and detect the Eml8 circulating antigen in patients' sera.


METHODS
Rabbits and BALB/c mice were immunized with the ReEml8 antigen, which was purified by affinity method for preparation of the specific poly-clonal and monoclonal antibodies. A sandwich ELISA was established by the specific antibodies.


RESULTS
By immunizing with the ReEml8 antigen, high antibody level was reached with a serum dilution of 1: 204 800 and above in the immunized rabbits. After double selection by ELISA using the ReEml8 antigen and block ELISA using both AE-positive and negative control sera, 14 positive cell clones were obtained with an inhibition rate of more than 50%. Those mono-and poly-clonal antibodies were matched freely in sandwich ELISA tests for detecting the ReEml8 antigen. A combination of monoclonal antibody No.9 and polyclonal antibody showed the best result. The sensitivity to detect ReEml8 antigen was at 3 ng/ml. Six of 11 AE sera were positive when tested with the sandwich ELISA system.


CONCLUSIONS
Highly specific polyclonal and monoclonal antibodies have been prepared, and a sensitive sandwich ELISA established. Preliminary result is suggested that a detectable level of Eml8 circulating antigen is present in AE patients' sera.
Jiang
Li
L
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai 200025, China.
Li
Xiong
X
Li
Hao
H
Niu
Xin-ling
XL
Feng
Zheng
Z
D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood*; D000947:Antigens, Helminth / Q000097:blood* / Q000235:genetics / Q000276:immunology; D004443:Echinococcosis / Q000097:blood* / Q000469:parasitology; D004444:Echinococcosis, Hepatic / Q000097:blood* / Q000469:parasitology; D048210:Echinococcus multilocularis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D011994:Recombinant Proteins