Blockade-of-Binding Activities toward Envelope-Associated, Type-Specific Epitopes as a Correlative Marker for Dengue Virus-Neutralizing Antibody

Abstract

This study describes a blockade-of-binding assay for the determination of antibodies that recognize a selected set of serotype-specific or group-reactive epitopes in the envelope of dengue virus. By employing blood samples collected from dengue virus-infected or -immunized macaques, moderate to strong correlations of the epitope-blocking activities with the virus-neutralizing antibody titers were observed with serotype-specific blocking activities for each of the four dengue serotypes.

Heterologous prime-boost immunization induces protection against dengue virus infection in cynomolgus macaques

“…8 ). A set of strongly neutralizing, type-specific monoclonal antibodies (1F4, 3H5, 8A1, and 5H2), which represent type-specific antibodies for DENV-1, DENV-2, DENV-3, and DENV-4, respectively, was shown with this assay to measure type-specific binding antibody activities with moderate or strong correlations with the corresponding neutralizing antibody titers in macaques ( 33 ). In agreement with the data from particle-binding experiments, the lowest relative level of 1F4-blocking activity at the time of challenge was detected in macaque ‘a’ ( Fig.…”

“…The blockade-of-binding assay was performed as described previously ( 33 ). Briefly, a pre-titrated amount of purified virus particles in PBS, pH 7.4, was applied to the wells of a 96-well microtiter plate (Maxisorp, Nunc) overnight at 4°C.…”

“…Moreover, the DENV-1 and -2 LAV components had been genetically modified by the introduction of a prM cleavage-enhancing mutation to increase the proportion of mature viral particles ( 29 – 31 ), which may more closely resemble the majority of circulating virions during dengue virus infection in humans ( 32 ). Antibody responses to immunization were determined by PRNT together with binding and blockade-of-binding assays to evaluate antibody properties, including the recognition of the whole viral particle, the receptor-binding domain of the major envelope protein E, and the E-associated serotype-specific epitopes ( 33 ).…”

Envelope-dimer epitope-like broadly protective antibodies against dengue in children following natural infection and vaccination

“…The correlation of EDE-like Abs with DENV2-specific neutralization was partly expected because EDE-like Abs were measured for binding to a DENV2 dimer. However, a recent animal study 61 s howed blockade of binding activity of EDE1 C10 was also moderately correlated with DENV2 monotypic neutralizing Abs 61 , suggesting DENV2 reactivity may be an intrinsic characteristic of EDE-like Abs. This suggests that some of the Abs, such as 2D22-like, that compete with EDE1 Abs in our assay may be specific to DENV2.…”

“…We developed the EDE BOB ELISA assay to measure the levels of human serum Abs that block the binding of EDE1 C8 mAb to the stabilized DENV2 E dimer. EDE BOB assays have been previously described 61,76 . Briefly, High binding 96-well flat-bottom microtiter plates (Microlon, Greiner, Germany) were coated with the capture anti-His mAb (Ref: TA150088, OriGene) in 1xTris-buffered saline (TBS) [50 mM tris-Cl (pH 7.5) and 150 mM NaCl] at 4°C overnight.…”