[Screening and expression of recombinant human monoclonal antibody Fab fragments specific to Entamoeba histolytica].
OBJECTIVE
To prepare recombinant human monoclonal antibody Fab fragments specific to the surface antigen of Entamoeba histolytica.


METHODS
Total RNA was isolated from lymphocytes which were separated from an asymptomatic E. histolytica cyst carrier. The genes of IgG light chain and Fd region of heavy chain were amplified by a reverse transcriptase PCR and ligated with a plasmid vector. After the genes were introduced into Escherichia coli, the clones expressing Fab fragments specific to the surface antigen of E. histolytica were screened and the product was purified.


RESULTS
Thirty thousand clones were screened and one of them was proved positive to the surface antigen of E. histolytica.


CONCLUSIONS
This study demonstrated that the bacterial system can be used to produce recombinant human monoclonal antibody Fab fragments specific to the surface antigen of E. histolytica and they may be applicable for the future diagnosis and treatment of the infection.
Department of Medical Microbiology & Parasitology, Shanghai Medical Center of Fudan University, Shanghai 200032, China.
Qing
Q
Shu
Hong-xia
HX
Sha
Hiroshi
H
Tachibana
Xun-jia
XJ
Cheng
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000913:Antibodies, Protozoan / Q000096:biosynthesis* / Q000235:genetics; D000953:Antigens, Protozoan / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D004748:Entamoeba histolytica / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins