Abstract
Varicella-zoster virus (VZV) is a highly contagious agent of varicella and herpes zoster. Varicella can be lethal to immunocompromised patients, babies, HIV patients and other adults with impaired immunity. Serological evaluation of immunity to VZV will help determine which individuals are susceptible and evaluate vaccine effectiveness. A collection of 110 monoclonal antibodies (mAbs) were obtained by immunization of mice with membrane proteins or cell-free virus. The mAbs were well characterized, and a competitive sandwich ELISA (capture mAb: 8H6; labelling mAb: 1B11) was established to determine neutralizing antibodies in human serum with reference to the FAMA test. A total of 920 human sera were evaluated. The competitive sandwich ELISA showed a sensitivity of 95.6%, specificity of 99.77% and coincidence of 97.61% compared with the fluorescent-antibody-to-membrane-antigen (FAMA) test. The capture mAb 8H6 was characterized as a specific mAb for VZV ORF9, a membrane-associated tegument protein that interacts with glycoprotein E (gE), glycoprotein B (gB) and glycoprotein C (gC). The labelling mAb 1B11 was characterized as a complement-dependent neutralizing mAb specific for the immune-dominant epitope located on gE, not on other VZV glycoproteins. The established competitive sandwich ELISA could be used as a rapid and high-throughput method for evaluating immunity to VZV.
Highlights
Varicella-zoster virus (VZV) is a highly contagious agent of varicella and herpes zoster
Labelling monoclonal antibodies (mAbs) (1B11) was characterized as a neutralizing mAb specific for the immune-dominant epitope located on glycoprotein E (gE)
The fragment of gN (21–49 aa) was fused to a linker (GSGGSG) and repeated four times; the new construct was named gN-L. gN-L was fused to a His tag and purified with Ni-NTA
Summary
Varicella-zoster virus (VZV) is a highly contagious agent of varicella and herpes zoster. The mAbs were well characterized, and a competitive sandwich ELISA (capture mAb: 8H6; labelling mAb: 1B11) was established to determine neutralizing antibodies in human serum with reference to the FAMA test. The competitive sandwich ELISA showed a sensitivity of 95.6%, specificity of 99.77% and coincidence of 97.61% compared with the fluorescent-antibody-to-membrane-antigen (FAMA) test. A highly sensitive and specific serological test is important for obtaining herd immunity against VZV. Several methods have been developed to measure serum VZV-specific immunoglobulin G (IgG) antibodies, including the fluorescent-antibody-to-membrane-antigen (FAMA) test[11,12], an enzyme-linked immunosorbent assay (ELISA) based on total VZV antigen or purified glycoproteins (gps)[13,14,15] and many other methods[10,16,17,18,19,20,21].
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