Biopanning of specific peptide for SARS-CoV-2 nucleocapsid protein and enzyme-linked immunosorbent assay-based antigen assay

Ma, Pengxin; Liu, Junchong; Pang, Shuang; Zhou, Wenhao; Yu, Haipeng; Wang, Mingyang; Dong, Tao; Li, Han; Wang, Qiqin; Liu, Aihua

doi:10.1016/j.aca.2023.341300

Cited by 8 publications

(5 citation statements)

References 66 publications

(68 reference statements)

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“…NP is essential for the early replication of the virus in the host cell, which plays a major role in packaging viral RNA into a ribonucleoprotein (RNP) complex called nucleocapsid. 325,326 When the virus enters the host cell, NP supports the replication of the viral RNA and releases the virus particles into the host cell. 327,328 However, because NP is highly conserved among all coronaviruses, the specificity of its detection is low.…”

Section: Sars-cov-2 Antigen Detectionmentioning

confidence: 99%

Diagnostics and analysis of SARS-CoV-2: current status, recent advances, challenges and perspectives

et al. 2023

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Section: Sars-cov-2 Antigen Detectionmentioning

confidence: 99%

Diagnostics and analysis of SARS-CoV-2: current status, recent advances, challenges and perspectives

et al. 2023

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“…(2) The biopanned phages are easy to prepare and purify, ready for scale-up application . Therefore, phage display has been widely used for virus-specific ligand screening, , disease diagnosis, − drug development, bioanalysis, and biosensing. , …”

Section: Introductionmentioning

confidence: 99%

Sensitive Dual-Signal ELISA Based on Specific Phage-Displayed Double Peptide Probes with Internal Filtering Effect to Assay Monkeypox Virus Antigen

Pang,

Wang,

Yuan

et al. 2024

Anal. Chem.

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The global spread of monkeypox has become a worldwide public healthcare issue. Therefore, there is an urgent need for accurate and sensitive detection methods to effectively control its spreading. Herein, we screened by phage display two peptides M4 (sequence: DPCGERICSIAL) and M6 (sequence: SCSSFLCSLKVG) with good affinity and specificity to monkeypox virus (MPXV) B21R protein. To simulate the state of the peptide in the phage and to avoid spatial obstacles of the peptide, GGGSK was added at the C terminus of M4 and named as M4a. Molecular docking shows that peptide M4a and peptide M6 are bound to different epitopes of B21R by hydrogen bonds and salt-bridge interactions, respectively. Then, peptide M4a was selected as the capture probe, phage M6 as the detection probe, and carbonized polymer dots (CPDs) as the fluorescent probe, and a colorimetric and fluorescent double-signal capture peptide/antigen/signal peptidedisplayed phage sandwich ELISA triggered by horseradish peroxidase (HRP) through a simple internal filtration effect (IFE) was constructed. HRP catalyzes H 2 O 2 to oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) to generate blue oxidized TMB, which can further quench the fluorescence of CPDs through IFE, enabling to detect MPXV B21R in colorimetric and fluorescent modes. The proposed simple immunoassay platform shows good sensitivity and reliability in MPXV B21R detection. The limit of detection for colorimetric and fluorescent modes was 27.8 and 9.14 pg/mL MPXV B21R, respectively. Thus, the established double-peptide sandwich-based dual-signal immunoassay provides guidance for the development of reliable and sensitive antigen detection capable of mutual confirmation, which also has great potential for exploring various analytical strategies for other respiratory virus surveillance.

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“…36,37 As the epidemic evolves and the virus mutates, the prevention and treatment of SARS-CoV-2 should be further strengthened to improve the accuracy and sensitivity of diagnostic methods. 38,39 Therefore, in this work, selecting SARS-CoV-2 as a proof of concept, we proposed a nonamplified bioscreening approach to identify peptides that can bind to the SARS-CoV-2 S protein with good affinity and specificity. Subsequently, we constructed the antibody-free pLFIA system using the identified peptide as a capture probe and developed a PDA@MnO 2 -based pLFIA with outstanding oxidase-like activity as a colorimetric tag for significant signal enhancement.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Polydopamine (PDA), possessing chemical and physical properties similar to those of natural melanin, along with strong adhesion and excellent biocompatibility, is considered to be a typical class of artificial melanin materials. − In addition, MnO 2 and MnO 2 -related materials exhibit prominent oxidase-like activities, which can catalyze 3,3′,5,5′-tetramethylbenzidine (TMB) oxidation with remarkable on-demand signal enhancement . The catastrophic spread of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused serious public healthcare problems and severely weakened the global economy. , As the epidemic evolves and the virus mutates, the prevention and treatment of SARS-CoV-2 should be further strengthened to improve the accuracy and sensitivity of diagnostic methods. , Therefore, in this work, selecting SARS-CoV-2 as a proof of concept, we proposed a nonamplified bioscreening approach to identify peptides that can bind to the SARS-CoV-2 S protein with good affinity and specificity. Subsequently, we constructed the antibody-free pLFIA system using the identified peptide as a capture probe and developed a PDA@MnO 2 -based pLFIA with outstanding oxidase-like activity as a colorimetric tag for significant signal enhancement.…”

Section: Introductionmentioning

confidence: 99%

Sensitive Lateral Flow Immunoassay Based on Specific Peptide and Superior Oxidase Mimics with a Universal Dual-Mode Significant Signal Amplification

et al. 2023

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Rapid and sensitive antigen detection using a lateral flow immunoassay (LFIA) is crucial for diagnosing infectious diseases due to its simplicity, speed, and user-friendly features. However, it remains a critical issue to explore specific biorecognition elements and powerful signal amplification. In this study, taking SARS-CoV-2 as a proof of concept, a specific peptide, WFLNDSELIML, binding to the SARS-CoV-2 spike (S) antigen was identified by a nonamplified biopanning method, which exhibited high affinity to the target, with a dissociation constant of 9.29 ± 1.55 nM. Molecular docking analysis reveals that this peptide binds to the N-terminal domain of the SARS-CoV-2 S antigen. Then, using this peptide as a capture probe and angiotensin-converting enzyme 2 as a detection probe, a peptide-based lateral flow immunoassay (pLFIA) for the sensitive detection of the SARS-CoV-2 S antigen without any antibody was developed, for which a polydopamine nanosphere (PDA)@MnO 2 nanocomposite with excellent oxidase-like activity was used as a colorimetric label, exhibiting dual-mode remarkable signal amplification of natural melanin and on-demand nanozyme catalytic enhancement. The PDA@MnO 2 -based pLFIA is capable of detecting the SARS-CoV-2 S antigen with a limit of detection of 8.01 pg/mL, which is 18.7 times lower than that of a conventional pLFIA tagged with gold nanoparticles. Additionally, the as-proposed PDA@MnO 2 -based pLFIA can detect up to 150 transduction units/mL SARS-CoV-2 pseudoviruses spiked in saliva samples. Given the outstanding analytical performance, the proposed PDA@MnO 2 -based pLFIA may offer a reliable option for the rapid diagnosis of SARS-CoV-2.

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Biopanning of specific peptide for SARS-CoV-2 nucleocapsid protein and enzyme-linked immunosorbent assay-based antigen assay

Cited by 8 publications

References 66 publications

Diagnostics and analysis of SARS-CoV-2: current status, recent advances, challenges and perspectives

Diagnostics and analysis of SARS-CoV-2: current status, recent advances, challenges and perspectives

Sensitive Dual-Signal ELISA Based on Specific Phage-Displayed Double Peptide Probes with Internal Filtering Effect to Assay Monkeypox Virus Antigen

Sensitive Lateral Flow Immunoassay Based on Specific Peptide and Superior Oxidase Mimics with a Universal Dual-Mode Significant Signal Amplification

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