Electrochemical Determination of Interaction between SARS-CoV-2 Spike Protein and Specific Antibodies

The serologic diagnosis of coronavirus disease 2019 (COVID-19) and the evaluation of vaccination effectiveness are identified by the presence of antibodies specific to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this paper, we present the electrochemical-based biosensing technique for the detection of antibodies specific to the SARS-CoV-2 proteins. Recombinant SARS-CoV-2 spike proteins (rSpike) were immobilised on the surface of a gold electrode modified by a self-assembled monolayer (SAM). This modified electrode was used as a sensitive element for the detection of polyclonal mouse antibodies against the rSpike (anti-rSpike). Electrochemical impedance spectroscopy (EIS) was used to observe the formation of immunocomplexes while cyclic voltammetry (CV) was used for additional analysis of the surface modifications. It was revealed that the impedimetric method and the elaborate experimental conditions are appropriate for the further development of electrochemical biosensors for the serological diagnosis of COVID-19 and/or the confirmation of successful vaccination against SARS-CoV-2

Characterization of monoclonal antibodies against hantavirus nucleocapsid protein and their use for immunohistochemistry on rodent and human samples

Fil: Kucinskaite-Kodze, Indre. Vilnius University. Institute of Biotechnology; Lituania.Fil: Petraityte-Burneikiene, Rasa. Vilnius University. Institute of Biotechnology; Lituania.Fil: Zvirbliene, Aurelija. Vilnius University. Institute of Biotechnology; Lituania.Fil: Hjelle, Brian. University of New Mexico School of Medicine. Department of Pathology; Estados Unidos.Fil: Medina, Rafael A. University of New Mexico School of Medicine. Department of Pathology; Estados Unidos.Fil: Gedvilaite, Alma. Vilnius University. Institute of Biotechnology; Lituania.Fil: Razanskiene, Ausra. Vilnius University. Institute of Biotechnology; Lituania.Fil: Schmidt-Chanasit, Jonas. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Fil: Mertens, Marc. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Fil: Padula, Paula. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Sasnauskas, Kestutis. Vilnius University. Institute of Biotechnology; Lituania.Fil: Ulrich, Rainer G. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Monoclonal antibodies are important tools for various applications in hantavirus diagnostics. Recently, we generated Puumala virus (PUUV)-reactive monoclonal antibodies (mAbs) by immunisation of mice with chimeric polyomavirus-derived virus-like particles (VLPs) harbouring the 120-amino-acid-long amino-terminal region of the PUUV nucleocapsid (N) protein. Here, we describe the generation of two mAbs by co-immunisation of mice with hexahistidine-tagged full-length N proteins of Sin Nombre virus (SNV) and Andes virus (ANDV), their characterization by different immunoassays and comparison with the previously generated mAbs raised against a segment of PUUV N protein inserted into VLPs. All of the mAbs reacted strongly in ELISA and western blot tests with the antigens used for immunization and cross-reacted to varying extents with N proteins of other hantaviruses. All mAbs raised against a segment of the PUUV N protein presented on chimeric VLPs and both mAbs raised against the full-length AND/SNV N protein reacted with Vero cells infected with different hantaviruses. The reactivity of mAbs with native viral nucleocapsids was also confirmed by their reactivity in immunohistochemistry assays with kidney tissue specimens from experimentally SNV-infected rodents and human heart tissue specimens from hantavirus cardiopulmonary syndrome patients. Therefore, the described mAbs represent useful tools for the immunodetection of hantavirus infection

Characterization of monoclonal antibodies against hantavirus nucleocapsid protein and their use for immunohistochemistry on rodent and human samples

Fil: Kucinskaite-Kodze, Indre. Vilnius University. Institute of Biotechnology; Lituania.Fil: Petraityte-Burneikiene, Rasa. Vilnius University. Institute of Biotechnology; Lituania.Fil: Zvirbliene, Aurelija. Vilnius University. Institute of Biotechnology; Lituania.Fil: Hjelle, Brian. University of New Mexico School of Medicine. Department of Pathology; Estados Unidos.Fil: Medina, Rafael A. University of New Mexico School of Medicine. Department of Pathology; Estados Unidos.Fil: Gedvilaite, Alma. Vilnius University. Institute of Biotechnology; Lituania.Fil: Razanskiene, Ausra. Vilnius University. Institute of Biotechnology; Lituania.Fil: Schmidt-Chanasit, Jonas. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Fil: Mertens, Marc. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Fil: Padula, Paula. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Sasnauskas, Kestutis. Vilnius University. Institute of Biotechnology; Lituania.Fil: Ulrich, Rainer G. Federal Research Institute for Animal Health. Institute of Epidemiology; Alemania.Monoclonal antibodies are important tools for various applications in hantavirus diagnostics. Recently, we generated Puumala virus (PUUV)-reactive monoclonal antibodies (mAbs) by immunisation of mice with chimeric polyomavirus-derived virus-like particles (VLPs) harbouring the 120-amino-acid-long amino-terminal region of the PUUV nucleocapsid (N) protein. Here, we describe the generation of two mAbs by co-immunisation of mice with hexahistidine-tagged full-length N proteins of Sin Nombre virus (SNV) and Andes virus (ANDV), their characterization by different immunoassays and comparison with the previously generated mAbs raised against a segment of PUUV N protein inserted into VLPs. All of the mAbs reacted strongly in ELISA and western blot tests with the antigens used for immunization and cross-reacted to varying extents with N proteins of other hantaviruses. All mAbs raised against a segment of the PUUV N protein presented on chimeric VLPs and both mAbs raised against the full-length AND/SNV N protein reacted with Vero cells infected with different hantaviruses. The reactivity of mAbs with native viral nucleocapsids was also confirmed by their reactivity in immunohistochemistry assays with kidney tissue specimens from experimentally SNV-infected rodents and human heart tissue specimens from hantavirus cardiopulmonary syndrome patients. Therefore, the described mAbs represent useful tools for the immunodetection of hantavirus infection