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2005•Zhongguo yaoke daxue xuebaoRequires access

Preparation and Identification of Monoclonal Antibody against Recombinant Human Erythropoietin

Tang Zu

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Abstract

AIM:To obtain monoclonal antibody (McAb) which can specificly identify human erythropoietin(hEPO).METHOD:Spleen cells from Balb/c mice immunized with recombinant human erythropoietin (rHuEPO) protien were fused with Sp2/0 myeloma cells.The specific McAb were screened by ELISA and identified by western blot.RESULT:Four hybridoma cell lines (3E_ 9,5A_ 6,4G_ 12and 2H_ 2) secreting specific McAb to human cytomegalo-virus (HCMV) were selected and determinded to recognize different epitops on gp52 protein.The titers of the four clones were as the follows:1∶2 048,1∶1 024,1∶1 024,1∶512(supernant);1×10 -7,1×10 -7,1×10 -6,2×10 -6 (AF).CONCLUSION:McAbs 2H_ 2 recognize one epitope on rhCMV gp52 protien,whereas 3E_ 9,5A_ 6and 4G_ 12 recognize another.

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AIM:To obtain monoclonal antibody (McAb) which can specificly identify human erythropoietin(hEPO).METHOD:Spleen cells from Balb/c mice immunized with recombinant human erythropoietin (rHuEPO) protien were fused with Sp2/0 myeloma cells.The specific McAb were screened by ELISA and identified by western blot.RESULT:Four hybridoma cell lines (3E_ 9,5A_ 6,4G_ 12and 2H_ 2) secreting specific McAb to human cytomegalo-virus (HCMV) were selected and determinded to recognize different epitops on gp52 protein.The titers of the four clones were as the follows:1∶2 048,1∶1 024,1∶1 024,1∶512(supernant);1×10 -7,1×10 -7,1×10 -6,2×10 -6 (AF).CONCLUSION:McAbs 2H_ 2 recognize one epitope on rhCMV gp52 protien,whereas 3E_ 9,5A_ 6and 4G_ 12 recognize another.

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Available abstract

AIM:To obtain monoclonal antibody (McAb) which can specificly identify human erythropoietin(hEPO).METHOD:Spleen cells from Balb/c mice immunized with recombinant human erythropoietin (rHuEPO) protien were fused with Sp2/0 myeloma cells.The specific McAb were screened by ELISA and identified by western blot.RESULT:Four hybridoma cell lines (3E_ 9,5A_ 6,4G_ 12and 2H_ 2) secreting specific McAb to human cytomegalo-virus (HCMV) were selected and determinded to recognize different epitops on gp52 protein.The titers of the four clones were as the follows:1∶2 048,1∶1 024,1∶1 024,1∶512(supernant);1×10 -7,1×10 -7,1×10 -6,2×10 -6 (AF).CONCLUSION:McAbs 2H_ 2 recognize one epitope on rhCMV gp52 protien,whereas 3E_ 9,5A_ 6and 4G_ 12 recognize another.

Key concepts: Erythropoietin, Monoclonal antibody, Recombinant DNA, Titer, Molecular biology, Epitope, Spleen, Western blot

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