Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests

Yamaoka, Yutaro; Miyakawa, Kei; Jeremiah, Sundararaj Stanleyraj; Funabashi, Rikako; Okihara, Koji; Kikuchi, Susumu; Katada, Junichi; Wada, Atsuhiko; Takei, Toshiki; Nishi, Mayuko; Shimizu, Kohei; Ozawa, Hiroki; Usuku, Shuzo; Kawakami, Chiharu; Tanaka, Nobuko; Morita, Takeshi; Hayashi, Hiroyuki; Mitsui, Hideaki; Suzuki, Komei; Aizawa, Daisuke; Yoshimura, Yukihiro; Miyazaki, Tomoyuki; Yamazaki, Etsuko; Suzuki, Tadaki; Kimura, Hirokazu; Shimizu, Hideaki; Okabe, Nobuhiko; Hasegawa, Hideki; Ryo, Akihide

doi:10.1016/j.xcrm.2021.100311

Cited by 41 publications

(63 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, they can be used for other applications, such as diagnosis and epidemiology. For example, monoclonal antibodies specifically targeting the SARS-CoV-2 N protein increase the detection sensitivity of ELISAs, providing a tool for the early and accurate diagnosis based on clinical samples and for epidemiological studies of SARS-CoV-2 [ 79 , 80 ]. In addition, the NP-monoclonal antibody system and related test strips can be utilized for large-scale screening of COVID-19 samples [ 81 ].…”

Section: Antigenic View Of the Ntd And Rbd Of Sars-cov-2 S Protein And Nab Binding Sitesmentioning

confidence: 99%

Neutralizing antibodies for the prevention and treatment of COVID-19

2021

View full text Add to dashboard Cite

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) initiates the infection process by binding to the viral cellular receptor angiotensin-converting enzyme 2 through the receptor-binding domain (RBD) in the S1 subunit of the viral spike (S) protein. This event is followed by virus–cell membrane fusion mediated by the S2 subunit, which allows virus entry into the host cell. Therefore, the SARS-CoV-2 S protein is a key therapeutic target, and prevention and treatment of coronavirus disease 2019 (COVID-19) have focused on the development of neutralizing monoclonal antibodies (nAbs) that target this protein. In this review, we summarize the nAbs targeting SARS-CoV-2 proteins that have been developed to date, with a focus on the N-terminal domain and RBD of the S protein. We also describe the roles that binding affinity, neutralizing activity, and protection provided by these nAbs play in the prevention and treatment of COVID-19 and discuss the potential to improve nAb efficiency against multiple SARS-CoV-2 variants. This review provides important information for the development of effective nAbs with broad-spectrum activity against current and future SARS-CoV-2 strains.

show abstract

Section: Antigenic View Of the Ntd And Rbd Of Sars-cov-2 S Protein And Nab Binding Sitesmentioning

confidence: 99%

Neutralizing antibodies for the prevention and treatment of COVID-19

2021

View full text Add to dashboard Cite

show abstract

“…The NP is a viral antigen that plays a crucial role in packaging viral genome RNA (Figure S2). The SARS-CoV-2 NP is abundantly expressed during the viral infection and is highly immunogenic [20][21][22]. Therefore, it is considered an ideal target for a diagnostic antigen of SARS-CoV-2 [21][22][23][24].…”

Section: Development Of Sars-cov-2 Np-specific Monoclonal Antibodies Derived From Short Conserved Peptides In the Np Antigenmentioning

confidence: 99%

“…The SARS-CoV-2 NP is abundantly expressed during the viral infection and is highly immunogenic [20][21][22]. Therefore, it is considered an ideal target for a diagnostic antigen of SARS-CoV-2 [21][22][23][24]. However, SARS-CoV-2 NP shares sequence homology with NPs from other coronaviruses (~90% identity with SARS-CoV NP and ~50% identity with MERS-CoV NP; Figure S1).…”

Section: Development Of Sars-cov-2 Np-specific Monoclonal Antibodies Derived From Short Conserved Peptides In the Np Antigenmentioning

confidence: 99%

Rapid Biosensor of SARS-CoV-2 Using Specific Monoclonal Antibodies Recognizing Conserved Nucleocapsid Protein Epitopes

Lee

Jung

Lee

et al. 2022

Viruses

View full text Add to dashboard Cite

Coronavirus disease 2019 (COVID-19), the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is characterized by symptoms such as fever, fatigue, a sore throat, diarrhea, and coughing. Although various new vaccines against COVID-19 have been developed, early diagnostics, isolation, and prevention remain important due to virus mutations resulting in rapid and high disease transmission. Amino acid substitutions in the major diagnostic target antigens of SARS-CoV-2 may lower the sensitivity for the detection of SARS-CoV-2. For this reason, we developed specific monoclonal antibodies that bind to epitope peptides as antigens for the rapid detection of SARS-CoV-2 NP. The binding affinity between antigenic peptides and monoclonal antibodies was investigated, and a sandwich pair for capture and detection was employed to develop a rapid biosensor for SARS-CoV-2 NP. The rapid biosensor, based on a monoclonal antibody pair binding to conserved epitopes of SARS-CoV-2 NP, detected cultured virus samples of SARS-CoV-2 (1.4 × 103 TCID50/reaction) and recombinant NP (1 ng/mL). Laboratory confirmation of the rapid biosensor was performed with clinical specimens (n = 16) from COVID-19 patients and other pathogens. The rapid biosensor consisting of a monoclonal antibody pair (75E12 for capture and the 54G6/54G10 combination for detection) binding to conserved epitopes of SARS-CoV-2 NP could assist in the detection of SARS-CoV-2 NP under the circumstance of spreading SARS-CoV-2 variants.

show abstract

“…With advantages on molecular stability, adjustable affinity, and batch consistency, aptamers are more suitable for developing low-cost sensors for large-scale applications. 28 , 29 To date, immunoassays for N-protein are mostly reported due to the availability of antibodies, 24 , 25 , 30 − 32 although it is encouraging that researchers have recently focused on aptasensor development. 27 , 33 Based on the traditional sensing methods, such as ELISA and electrochemistry, the lowest limit of detection (LOD) of the abovementioned assays is 6.25 pg/mL.…”

Section: Introductionmentioning

confidence: 99%

“… 27 , 33 Based on the traditional sensing methods, such as ELISA and electrochemistry, the lowest limit of detection (LOD) of the abovementioned assays is 6.25 pg/mL. 32 At the same time, the shortest turnaround time is not satisfied (about 1 h) due to the lack of efficient techniques for target enrichment. 25 …”

Section: Introductionmentioning

confidence: 99%

Capacitive Aptasensor Coupled with Microfluidic Enrichment for Real-Time Detection of Trace SARS-CoV-2 Nucleocapsid Protein

Yang

et al. 2022

Anal. Chem.

View full text Add to dashboard Cite

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has lasted for almost 2 years. Stemming its spread has posed severe challenges for clinical virus detection. A long turnaround time, complicated operation, and low accuracy have become bottlenecks in developing detection techniques. Adopting a direct antigen detection strategy, we developed a fast-responding and quantitative capacitive aptasensor for ultratrace nucleocapsid protein detection based on a low-cost microelectrode array (MEA) chip. Employing the solid–liquid interface capacitance with a sensitivity of picofarad level, the tiny change on the MEA surface can be definitively detected. As a result, the limit of detection reaches an ultralow level of femtogram per milliliter in different matrices. Integrated with efficient microfluidic enrichment, the response time of this sensor from the sample to the result is shortened to 15 s, completely meeting the real-time detection demand. Moreover, the wide linear range of the sensor is from 10 –5 to 10 –2 ng/mL, and a high selectivity of 6369:1 is achieved. After application and evaluation in different environmental and body fluid matrices, this sensor and the detection method have proved to be a label-free, real-time, easy-to-operate, and specific strategy for SARS-CoV-2 screening and diagnosis.

show abstract

Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests

Cited by 41 publications

References 54 publications

Neutralizing antibodies for the prevention and treatment of COVID-19

Neutralizing antibodies for the prevention and treatment of COVID-19

Rapid Biosensor of SARS-CoV-2 Using Specific Monoclonal Antibodies Recognizing Conserved Nucleocapsid Protein Epitopes

Capacitive Aptasensor Coupled with Microfluidic Enrichment for Real-Time Detection of Trace SARS-CoV-2 Nucleocapsid Protein

Contact Info

Product

Resources

About