2003•Journal of Preventive Medicine of the Chinese People's Liberation ArmyRequires access
PREPARATION AND APPLICATION OF MONOCLONAL ANTIBODY AGAINST RECOMBINANT HUMAN CYTOMEGALOVIRUS gp52 PROTEIN
Zhou Zong-an
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Abstract
Objective:To obtain monoclonal antibody (McAb) against recombinant human cytomegalovirus (rhCMV) for the detection of human cytomegalovirus (HCMV) IgM antibody following infection. Methods:McAbs against rhCMV gp52 protein were obtained by hybridoma technique.Antibody-capture ELISA (AC-ELISA) were composed of coated antibody (anti-human μ chain),antigen (rhCMV gp52)and marker antibody (specific HRP-McAbs). 939 sera were examined for CMV-IgM by AC-ELISA. Results: Four hybridoma cell lines (3E9,5A6,4G12 and 2H2) secreting high specific McAbs against rhHCMV were obtained, which recognized 2 different epitopes on rhCMV gp52 protein. 2H2 recognized one epitope while 3E9,5A6, and 4G12 recognized together the other one.The proper content of gp52 was 1μg·mL -1 and working dilution of HRP-McAbs was 1:1 600. Within-run CV was 3.7%, between-run CV was 7.9%. 17 of 643 (2.7%) blood donors, 10 of 288 (3.5%) patients with virus hepatitis and 11 of 17(64.7%) patients with hepatitis syndrome developed anti-HCMV IgM. Conclusion:The AC-ELISA using recombinant gp52 antigen and McAbs is specific, sensitive, not interfered by rheumatoid factor and reliable for the detection of serum HCMV-IgM.It is better than CMV antigen and rabbit antibody ELISA and is suitable for clinical diagnosis and epidemiological investigation.