Nanobody-Based Blocking of Binding ELISA for the Detection of Anti-NS1 Zika-Virus-Specific Antibodies in Convalescent Patients

• Published in: Tropical medicine and infectious disease Vol. 8; no. 1; p. 55
• Main Authors: Delfin-Riela, Triana, Rossotti, Martín A, Mattiuzzo, Giada, Echaides, César, González-Sapienza, Gualberto
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 10.01.2023; MDPI
• Subjects: Antibodies; Antigens; Biomarkers; Dengue fever; Enzymes; flavivirus; immunoassay; Infections; phage display; Proteins; Serology; single-domain antibodies; Zika virus; United Kingdom > UK; United States > US; single-domain antibodies; phage display; serology; flavivirus; immunoassay
• ISSN: 2414-6366; 2414-6366
• DOI: 10.3390/tropicalmed8010055

Zika virus has spread around the world with rapid pace in the last five years. Although symptoms are typically mild and unspecific, Zika's major impact occurs during pregnancy, generating a congenital syndrome. Serology plays a key role in its diagnosis. However, its use is limited due to the uncertainty caused by the cross-reaction of antibodies elicited in response to other flavivirus infections when tested in direct immunoassays. Using a panel of previously generated anti-Zika non-structural protein 1 (NS1) nanobodies, a set was selected that only recognizes epitopes present in Zika and is immunogenic to humans. A proper arrangement of these nanobodies was made and conditions were optimized in order to develop a novel serology assay. This new ELISA relies on the inhibition of the binding of a set of selected nanobodies to Zika-immobilized NS1 when previously incubated with Zika convalescent sera. Using the developed blocking of binding assay, it was possible to discriminate between Zika-specific and cross-reactive antibodies in serum samples from infections with Zika and other flaviviruses.