Broad-spectrum monoclonal antibodies against chikungunya virus structural proteins: Promising candidates for antibody-based rapid diagnostic test development

Abstract

In response to the aggressive global spread of the mosquito-borne chikungunya virus (CHIKV), an accurate and accessible diagnostic tool is of high importance. CHIKV, an arthritogenic alphavirus, comprises three genotypes: East/Central/South African (ECSA), West African (WA), and Asian. A previous rapid immunochromatographic (IC) test detecting CHIKV E1 protein showed promising performance for detection of the ECSA genotype. Unfortunately, this kit exhibited lower capacity for detection of the Asian genotype, currently in circulation in the Americas, reflecting the low avidity of one of the monoclonal antibodies (mAbs) in this IC kit for the E1 protein of the Asian-genotype because of a variant amino acid sequence. To address this shortcoming, we set out to generate a new panel of broad-spectrum mouse anti-CHIKV mAbs using hybridoma technology. We report here the successful generation of mouse anti-CHIKV mAbs targeting CHIKV E1 and capsid proteins. These mAbs possessed broad reactivity to all three CHIKV genotypes, while most of the mAbs lacked cross-reactivity towards Sindbis, dengue, and Zika viruses. Two of the mAbs also lacked cross-reactivity towards other alphaviruses, including O'nyong-nyong, Ross River, Mayaro, Western Equine Encephalitis, Eastern Equine Encephalitis, and Venezuelan Equine Encephalitis viruses. In addition, another two mAbs cross-reacted weakly only with most closely related O'nyong-nyong virus. Effective diagnosis is one of the keys to disease control but to date, no antibody-based rapid IC platform for CHIKV is commercially available. Thus, the application of the mAbs characterized here in the rapid diagnostic IC kit for CHIKV detection is expected to be of great value for clinical diagnosis and surveillance purposes.

Assessment of Immunogenicity and Neutralisation Efficacy of Viral-Vectored Vaccines Against Chikungunya Virus

“…In cells transduced with ChAdOx1 sCHIKV ∆C we also detected bands at approximately 55 kDa in supernatants (Figure 3c, lane 5,6,7), but no bands of 50 kDa or 18 kDa were found in the concentrated supernatant nor in the cell extracts. Therefore, it is suggested that the 35 kDa or 18 kDa band may represent the capsid protein, as previously demonstrated [38], which are not present in ChAdOx1 sCHIKV ∆C-transduced cells. As a positive control, we used a commercially available CHIKV E1 protein (Figure 3c, lane 9); therefore, it is suggested that the 55 kDa band detected in our blots is E1 protein.…”

Arthritogenic Alphaviruses: A Worldwide Emerging Threat?

“…Hence, the closer alphaviruses are phylogenetically, the more cross-reactivity exists [194]. In response, considerable effort is made on developing technologies to avoid cross-reactivity both in direct [271,277] and indirect diagnosis [194,278].…”

Antiviral therapeutics for chikungunya virus

“…The reactivity was reported against East/Central/South African (ECSA), West African, and Asian genotypes and MAbs also possessed no cross reactivity. Moreover, these MAbs are being used to detect E1 glycoprotein for detection of CHIKV infection via rapid immunochromatographic (IC) detection kits and worked well for all three CHIKV genotypes and possess great value in clinical diagnosis [80].…”

“…MAb targeting the E1 glycoprotein of CHIKV i.e. CK47 restricted the viral dissemination via inhibiting its budding during replication [80]. Fox et al reported the importance of interaction between Fc region of therapeutic MAbs and Fcγ receptors (FcγR) of effector cells such as monocytes during CHIKV infection.…”