Fabrication of a Lateral Flow Assay for Rapid In-Field Detection of COVID-19 Antibodies Using Additive Manufacturing Printing Technologies

Alrashoudi, Abdulelah A.; Albalawi, Hamed I.; Aldoukhi, Ali H.; Moretti, Manola; Bilalis, Panayiotis; Abedalthagafi, Malak; Hauser, Charlotte

doi:10.18063/ijb.v7i4.399

Cited by 11 publications

(7 citation statements)

References 29 publications

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“…In these assays, there is still a need to find an optimal antigen for better antibody detection, as recently highlighted by Rosadas et al Furthermore, these peptide epitopes could be used to develop rapid serological tests to detect SARS-CoV-2 antibodies. For example, a lateral flow immunoassay (LFIA) could be developed for the rapid detection of SARS-CoV-2 antibodies. , In addition, conjugating peptide epitopes to gold nanoparticles could enable rapid detection of antibodies using a nanosensor colorimetric assay . These rapid tests could be used as point-of-care tests where access to a clinical laboratory is limited.…”

Section: Resultsmentioning

confidence: 99%

“…For example, a lateral flow immunoassay (LFIA) could be developed for the rapid detection of SARS-CoV-2 antibodies. 41,42 In addition, conjugating peptide epitopes to gold nanoparticles could enable rapid detection of antibodies using a nanosensor colorimetric assay. 43 These rapid tests could be used as pointof-care tests where access to a clinical laboratory is limited.…”

Section: Future Directionsmentioning

confidence: 99%

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Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Aldoukhi,

Bilalis,

Alhattab

et al. 2023

ACS Bio Med Chem Au

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The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

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Section: Resultsmentioning

confidence: 99%

Section: Future Directionsmentioning

confidence: 99%

Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Aldoukhi,

Bilalis,

Alhattab

et al. 2023

ACS Bio Med Chem Au

Self Cite

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“…In addition, the rapid development of microscale additive manufacturing techniques could potentially further increase the detection sensitivity by incorporating multi-scale hierarchical structures. 31,32…”

Section: Discussionmentioning

confidence: 99%

Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

Bao,

Dollery,

Yuqing

et al. 2024

Lab Chip

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“…A needle was placed onto the microfluidic pump for dispensing. 28 Moreover, a low-cost, open-source, 3D-printed antibody dispenser was manufactured for line construction that consisted of common electronics (arduino, motors, and an LCD panel), two wheels, and a syringe needle. The membrane was moved by two wheels in a track, and the reagent was passively dispensed upon contact of the needle with the membrane surface.…”

Section: ■ Introductionmentioning

confidence: 99%

“…An in-house robotic arm and microfluidic pumps were utilized to dispense antibodies onto the membrane to detect SARS-CoV-2. A needle was placed onto the microfluidic pump for dispensing . Moreover, a low-cost, open-source, 3D-printed antibody dispenser was manufactured for line construction that consisted of common electronics (arduino, motors, and an LCD panel), two wheels, and a syringe needle.…”

Section: Introductionmentioning

confidence: 99%

Design and Validation of a Three-Dimensional Printer-Based System Enabling Rapid, Low-Cost Construction of the Biosensing Areas of Lateral Flow Devices for Immunoassays and Nucleic Acid Assays

Kalligosfyri,

Tragoulias,

Tsikas

et al. 2023

Anal. Chem.

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The COVID-19 pandemic proved the great usefulness of lateral flow tests as self- and rapid tests. The rapid expansion of this field requires the design and validation of novel, affordable, and versatile technologies for the easy fabrication of a variety of lateral flow devices. In the present work, we have developed a new, simple, and cost-effective system for the dispensing of reagents on the membranes of lateral flow devices to be used for research purposes. The 3D printing technology is integrated, for the first time, with simple and inexpensive tools such as a technical pen and disposable pipet tips for the construction of the test and the control areas of the devices. We also used this system for the automated fabrication of spots on the membrane for multiplex analysis. The devices were applied for the detection of proteins/antibodies and single- and double-stranded DNA targets. Also, devices with multiple biosensing areas on the membrane were constructed for the simultaneous detection of different analytes. The proposed system is very simple, automated, and inexpensive and has provided rapid and reproducible construction of lateral flow devices. Compared to a commercially available automated dispenser, the devices showed similar detection capabilities and reproducibility in various real samples. Moreover, contrary to the existing dispensers, the proposed system does not require any gas or costly precision pumps and syringes for the deposition. In conclusion, the developed 3D printer-based system could be an extremely useful alternative for research laboratories for the construction of lateral flow devices of various assay configurations.

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Fabrication of a Lateral Flow Assay for Rapid In-Field Detection of COVID-19 Antibodies Using Additive Manufacturing Printing Technologies

Cited by 11 publications

References 29 publications

Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

Design and Validation of a Three-Dimensional Printer-Based System Enabling Rapid, Low-Cost Construction of the Biosensing Areas of Lateral Flow Devices for Immunoassays and Nucleic Acid Assays

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