A colorimetric lateral flow immunoassay based on oriented antibody immobilization for sensitive detection of SARS-CoV-2

Lee, Ae Sol; Kim, Sumin; Kim, Kyeong Rok; Park, Chulmin; Lee, Dong Gun; Heo, Hye Ryoung; Joon, Hyung; Kim, Chang Sup

doi:10.1016/j.snb.2022.133245

Cited by 28 publications

(25 citation statements)

References 37 publications

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“…6A). 29 The developed LFA strip enabled sensitive detection of cultured SARS-CoV-2 in 15 min with a LoD of 5 × 10 4 copies per mL. The accuracy rate was verified to be 100% for 19 clinical positive and negative samples using RT-PCR.…”

Section: Poct Devices For Covid-19mentioning

confidence: 88%

Recent advances in point-of-care testing of COVID-19

Lee,

Bi,

Chen

et al. 2023

Chem. Soc. Rev.

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Section: Poct Devices For Covid-19mentioning

confidence: 88%

Recent advances in point-of-care testing of COVID-19

Lee,

Bi,

Chen

et al. 2023

Chem. Soc. Rev.

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“…Erroneous positive and negative molecular diagnostic results are indeed important and can have serious implications in people. Therefore, the use of RT-PCR combined with immunodiagnostic tests, such as LFIA or ELISA (enzyme-linked immunosorbent assay), can enhance the overall sensitivity and reduce erroneous information [44][45][46][47][48].…”

Section: Discussionmentioning

confidence: 99%

A Case Report of a COVID-19 Infection with Positive Sputum and Negative Nasopharyngeal Rapid Antigen-Based Testing: A Practical Method to Explore Sputome

Guzman¹,

Guzman

2023

Preprint

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Current COVID19 antigen testing is primarily carried out by obtaining a specimen via nasopharyngeal swab and performing a rapid lateral flow immunoassay, LFIA, or related immunoassays. On average, a nasopharyngeal antigen based LFIA for COVID19 remains positive for approximately one week from symptom onset, and levels of infectivity and duration of the symptoms may depend primarily on carrying a high viral load enough to infect others. It has been proposed that patients with long COVID, a syndrome in which patients continue to have complications of COVID with ongoing symptoms, may have occurring viral replication, despite testing negative via rapid COVID antigen testing. We therefore propose a modified antigen-based method that exposes hidden or masked antigenic sites of viral specimens, or lingering fragments of viral proteins, present in sputum using a home based rapid immunoassay for COVID19. Almost all protocols for testing were performed according to LFIA kit manufacturer s instructions for the detection of SARSCoV2 nucleocapsid protein antigen, one of the most predominant proteins encoded by the SARSCoV2 virus. However, in challenging the manufacturer s instructions, one or more digestive enzymes and a detergent were added to the collected biosamples (nasopharyngeal, oropharyngeal, saliva, buccal, gargle, and sputum); this modified procedure expose hidden or masked antigenic sites of the coronavirus or crossreactant antigenic sites of related or non-related viruses, or some of the plethora of epitopes generated by the sample corresponding microbiota to accomplish an optimal binding to the commercial antibody used in the diagnostic test. The modified protocol can enhance detection sensitivity by making the resultant test band in sputum samples visible, that would otherwise not be seen, and consequently may generate a false negative result, in a nasopharyngeal sample from a patient with mild symptoms of COVID19 and or low viral load. Therefore, a need exists for an improved sample pre-treatment extraction procedure that allows optimization of sample preparation to attain a more accurate test result. Although the experiments described here were performed using commercial platforms, with antibodies directed to SARSCoV2 nucleocapsid antigen, this method may also be viable for the detection of any other pathogen in sputum by using antibodies directed to the key antigens present in the pathogen of interest. Furthermore, this modified method to expose the content of sputum can be used as a simple protocol to study the sputome, the proteome of sputum, and other omics, sputomics. In summary, this simple method is non-invasive, rapid, inexpensive, accurate, and may provide increased sensitivity and specificity in the detection of COVID19 antigens for several weeks or even months

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“…For instance, in antigen testing, the sample migrates to the absorption pad under the influence of capillary action. If the analyte is present, then the labeled capture antibody binds to the detection antibody through the analyte, producing a positive outcome; otherwise, the result will be negative. , Notably, the LFIA method primarily relies on monoclonal antibodies (mAbs) comprising two heavy chains and two light chains . Conventional antibodies are generated through hybridoma technology, necessitating a high production cost and a protracted production timeline.…”

Section: Introductionmentioning

confidence: 99%

“…If the analyte is present, then the labeled capture antibody binds to the detection antibody through the analyte, producing a positive outcome; otherwise, the result will be negative. 22,23 Notably, the LFIA method primarily relies on monoclonal antibodies (mAbs) comprising two heavy chains and two light chains. 24 production timeline.…”

Section: ■ Introductionmentioning

confidence: 99%

Nanobody in a Double “Y”-Shaped Assembly: A Promising Candidate for Lateral Flow Immunoassays

Xiao,

Dong,

et al. 2024

Anal. Chem.

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Derived from camelid heavy-chain antibodies, nanobodies (Nbs) are the smallest natural antibodies and are an ideal tool in biological studies because of their simple structure, high yield, and low cost. Nbs possess significant potential for developing highly specific and user-friendly diagnostic assays. Despite offering considerable advantages in detection applications, knowledge is limited regarding the exclusive use of Nbs in lateral flow immunoassay (LFIA) detection. Herein, we present a novel double "Y" architecture, achieved by using the SpyTag/SpyCatcher and Im7/CL7 systems. The double "Y" assemblies exhibited a significantly higher affinity for their epitopes, as particularly evident in the reduced dissociation rate. An LFIA employing double "Y" assemblies was effectively used to detect the severe acute respiratory syndrome coronavirus-2 N protein, with a detection limit of at least 500 pg/mL. This study helps broaden the array of tools available for the development of Nb-based diagnostic techniques.

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A colorimetric lateral flow immunoassay based on oriented antibody immobilization for sensitive detection of SARS-CoV-2

Cited by 28 publications

References 37 publications

Recent advances in point-of-care testing of COVID-19

Recent advances in point-of-care testing of COVID-19

A Case Report of a COVID-19 Infection with Positive Sputum and Negative Nasopharyngeal Rapid Antigen-Based Testing: A Practical Method to Explore Sputome

Nanobody in a Double “Y”-Shaped Assembly: A Promising Candidate for Lateral Flow Immunoassays

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