Production of monoclonal antibodies using an electrophoretically separated diagnostic marker from the urine of individuals infected with Schistosoma japonicum.
In this paper, electrophoresis was successfully used to screen a diagnostic marker (a glycoprotein of 30 kDa) from the urine of individuals infected with Schistosoma japonicum, and then with the marker as antigen, two lines of monoclonal antibodies (mAbs) were obtained, NP56 and NP54. NP56 was of better immunoreactivity, its immunoglobulin isotype being IgG2b. NP56 reacted with soluble egg antigen and adult worm antigen and miracidia in eggs of S. japonicum. Using NP56 as a probe in indirect ELISA, the sensitivity and specificity (median egg excretion per gram of feces 69) was 90% and 100% in concentrated urine samples and 50% and 100% in original urine samples, respectively. The results indicate that the method is feasible for producing mAbs with diagnostic markers separated electrophoretically from the urine of individuals infected with pathogen.
Changliang
Zhu
Z
Department of Parasitology, Nanjing Medical University, People's Republic of China. clzhu@njmu.edu.cn
Haisheng
Tian
T
Lin
Lu
L
Yunqian
Li
L
Xiaohong
Guan
G
D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000947:Antigens, Helminth / Q000737:chemistry / Q000276:immunology; D015415:Biomarkers / Q000652:urine; D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D006801:Humans; D007150:Immunohistochemistry; D008970:Molecular Weight; D012549:Schistosoma japonicum / Q000254:growth & development / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000175:diagnosis* / Q000523:psychology / Q000652:urine; D012680:Sensitivity and Specificity
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D015415:Biomarkers