Performance Of Lateral Flow Immunoassay Research Articles

NIR-II fluorescence lateral flow immunosensor based on efficient energy transfer probe for point-of-care testing of tumor biomarkers

Fluorescence lateral flow immunoassay (LFA) has emerged as a powerful tool for rapid screening of various biomarkers owing to its simplicity, sensitivity and flexibility. It is noteworthy that fluorescent probe mainly determines the analytical performance of LFA. Due to the emission and excitation wavelengths are located in the visible region, most fluorophores are inevitably subject to light scattering and background autofluorescence. Herein, we reported a novel LFA sensor based on the second near-infrared (NIR-II) fluorescent probe with excellent anti-interference capability. The designed NIR-II probe was the Nd3+ and Yb3+ doped rare earth nanoparticles (RENPs) by employing Nd3+ as energy donor and Yb3+ as energy acceptor, which of the donor-acceptor energy transfer (ET) efficiency reached up to 80.7 %. Meanwhile, relying on the convenient and effective encapsulation strategy of poly(lactic-co-glycolic acid) (PLGA) microspheres to RENPs, the surface functionalized NIR-II probe (RE@PLGA) was obtained for subsequent bioconjugation. Benefiting from the optical advantages of NIR-II probe, this proposed NIR-II LFA displayed a good linear relationship ranging from 7 ng/mL to 200 ng/mL for the detection of α-fetoprotein (AFP), an important biomarker of hepatocellular carcinoma (HCC). The limit of detection (LOD) was determined as low as 3.0 ng/mL, which was of 8.3 times lower than clinical cutoff value. It is promising that LFA sensor based on this efficient RENPs probe provides new opportunities for high sensitive detection of various biomarkers in biological samples.

Goat anti-mouse immunoglobulin as “crosslinker” assisted signal tracer assemble with intensive antibody utilization efficiency for sensitive paper-based strip nanobiosensors

Engineered collaborative biochemical techniques and regulated nanomaterials (NMs) offer extraordinary opportunities for improving the analysis performance of lateral flow immunoassay (LFIA). Herein, inspired by the ability of macromolecules (e.g., proteins) to assemble into new functional units and the remarkable optical performance of engineered regulated NMs, goat anti-mouse immunoglobulin (GAMI) serves as the “crosslinker” integrate with gold‑manganese oxide (Au-MnOx) to assemble the “signal tracers (STs)-crosslinker-antibody (mAb)” for elevating the mAb utilization efficiency. Notably, the “STs-crosslinker-mAb” assembly shows ~13.33-folds mAb utilization efficiency enhance, which perfectly response the challenge between limited sensitivity and sufficient signal intensity in competitive-type LFIA. The black color and rough structure of Au-MnOx offer higher colorimetric brightness (~2-folds than AuNPs) and enhanced mAb coupling efficiency (up to 92.47%), which further improves sensitivity under the premise of functional assembly to intensify the competitive immunoreaction. Additionally, the convenient synthesis conditions (~13 min at room temperature) even comparable to direct purchase commercial products indicate that using Au-MnOx undoubtedly increases the cost-effectiveness. Encouragingly, the Au-MnOx-GAMI-mAb based LFIA exhibited high sensitivity (LOD: 0.063 ng mL−1 for clenbuterol (CLE) monitoring) by elevating mAb utilization efficiency with the attendant enhancing immune competition response in a cost-effective manner, which provides an invigorating reference pathway in point-of-care immunoassay.

Pt@AuNF nanozyme and horseradish peroxidase-based lateral flow immunoassay dual enzymes signal amplification strategy for sensitive detection of zearalenone

Lateral flow immunoassay (LFIA) has been employed extensively for the rapid, accurate, and portable detection of foodborne toxins. Here, the platinum gold nanoflower core-shell (Pt@AuNF) nanozyme with excellent optical properties, good catalytic ability and controllable reaction conditions were prepared to effectively improve the performance of lateral flow immunoassay (LFIA) strips. The Pt@AuNF nanozyme and horseradish peroxidase (HRP) combined with monoclonal antibody were used as signal probes based on the dual enzymes catalytic signal amplification strategy to detect Zearalenone sensitively. Dual enzymes catalyze the decomposition of hydrogen peroxide into hydroxyl radicals, and under the influence of hydroxyl radicals, colorless 3,3′,5,5′ -tetramethylbenzidine (TMB) is oxidized to blue ox-TMB, which is superimposed on the strips for signal amplification to broaden the detection range. The limit of detection (LOD) of the Pt@AuNF-HRP labeled LFIA strips after signal amplification was 0.052 ng/mL, and the detection range was 0.052–7.21 ng/mL. Compared with the Pt@AuNF labeled strips, while reducing the probes amount by half to achieve antibody conservation, the detection range was expanded by 5-fold based on achieving improved sensitivity. The study provided a meaningful reference for expanding the detection range based on immunoassay.

Quantum dots’ size matters for balancing their quantity and quality in label materials to improve lateral flow immunoassay performance for C-reactive protein determination

Incorporating quantum dots (QDs) into dendritic mesoporous silica nanoparticles (DMSNs) for signal amplification of label materials represents an efficient strategy to improve the performance of lateral flow immunoassays (LFIAs). In this work, it is found that the CdSe/ZnS QD's size matters for balancing their loading amount and quantum yields (QYs) in the DMSNs-QDs based label materials and ultimately determining the performance of LFIA. The impacts of three CdSe/ZnS QDs with diameters of 9.1, 10.5 and 11.7 nm on CdSe/ZnS QDs incorporation and LFIA applications are studied. The increase of CdSe/ZnS QDs size from 9.1 to 11.7 nm results in a decrease in CdSe/ZnS QDs loading amount and an increase in QYs of incorporated CdSe/ZnS QDs. This trade-off leads to an optimized CdSe/ZnS QDs size of 10.5 nm, which exhibits the best LFIA performance due to the balanced QDs loading (2.26 g g−1) and QY (57.1%). The 10.5 nm CdSe/ZnS QDs incorporated DMSNs-QDs for C-reactive protein (CRP) detection achieved a limit of detection of 5 pg mL−1 (equivalent to 4.2 × 10−14 M) with naked eye, which is lower than literature reports and commercial LFIA products. This study demonstrates that the CdSe/ZnS QD's size matters for improving the quality of DMSNs-QDs and their LFIA performance for CRP determination, providing new insights into the rational design of advanced label materials for improving LFIA performance.

Sensitive and quantitative detection of cardiac troponin I with upconverting nanoparticle lateral flow test with minimized interference

Measurement of cardiac troponin I (cTnI) should be feasible for point-of-care testing (POCT) to diagnose acute myocardial infarction (AMI). Lateral flow immunoassays (LFIAs) have been long implemented in POCT and clinical settings. However, sensitivity, matrix effect and quantitation in lateral flow immunoassays (LFIAs) have been major limiting factors. The performance of LFIAs can be improved with upconverting nanoparticle (UCNP) reporters. Here we report a new methodological approach to quantify cTnI using UCNP-LFIA technology with minimized plasma interference. The performance of the developed UCNP-LFIA was evaluated using clinical plasma samples (n = 262). The developed UCNP-LFIA was compared to two reference assays, the Siemens Advia Centaur assay and an in-house well-based cTnI assay. By introducing an anti-IgM scrub line and dried EDTA in the LFIA strip, the detection of cTnI in plasma samples was fully recovered. The UCNP-LFIA was able to quantify cTnI concentrations in patient samples within the range of 30–10,000 ng/L. The LoB and LoD of the UCNP-LFIA were 8.4 ng/L and 30 ng/L. The method comparisons showed good correlation (Spearman’s correlation 0.956 and 0.949, p < 0.0001). The developed UCNP-LFIA had LoD suitable for ruling in AMI in patients with elevated cTnI levels and was able to quantify cTnI concentrations in patient samples. The technology has potential to provide simple and rapid assay for POCT in ED setting

Boronate affinity-assisted oriented antibody conjugation on quantum dot nanobeads for improved detection performance in lateral flow immunoassay

The biological activity of immunoprobes is one of the most factors to determine the analytical performance of lateral flow immunoassay. However, the immunoprobes prepared using the carbodiimide method experience some limitations, such as random and disorder antibody orientation, thus reducing their binding affinity against the target antigen. Herein, we reported a facile approach for one-pot fabrication of boronate-tagged quantum dot nanobeads (QBs), and the boronate affinity reaction was introduced to assist the antibody-oriented attachment on the surface of QBs by the specific reaction of the boric acid group on QB surface and the carbohydrate residues containing cis-diol on the Fc fragment of antibodies. The fabricated immunoprobes were then used for the LFIA of hepatitis B surface antigen at the optimal conditions. In comparison with the carbodiimide method, the proposed conjugation strategy showed better antibody orientation, lower usage of labeled antibody (25 µg antibodies per mg boronate-tagged QBs vs. 350 µg antibodies per mg carboxyl modified QBs), shorter coupling time (5 min vs. 90 min), and higher antibody biological activity (10.8-fold higher than that of carboxyl-modified QBs). Furthermore, the developed boronate-tagged QB-based LFIA exhibited a low detection limit of 0.062 ng mL−1 and a wide dynamic linear range from 0.1 ng mL−1 to 1600 ng mL−1 for the hepatitis B surface antigen quantitativedetermination. In addition, the proposed oriented antibody strategy also showed the general applicability for detecting other four biomarkers by simply replacing the corresponding antibodies. In brief, this method is promising as a universal fabrication technology for the preparation of high-performance nanoparticle assemblies-based immunoprobes with directional antibody conjugation.

Analytical performance of lateral flow immunoassay for SARS-CoV-2 exposure screening on venous and capillary blood samples

ObjectivesWe validate the use of a lateral flow immunoassay (LFI) intended for rapid screening and qualitative detection of anti-SARS-CoV-2 IgM and IgG in serum, plasma, and whole blood, and compare results with ELISA. We also seek to establish the value of LFI testing on blood obtained from a capillary blood sample. MethodsSamples collected by venous blood draw and finger stick were obtained from patients with SARS-CoV-2 detected by RT-qPCR and control patients. Samples were tested with Biolidics 2019-nCoV IgG/IgM Detection Kit lateral flow immunoassay, and antibody calls were compared with ELISA. ResultsBiolidics LFI showed clinical sensitivity of 92% with venous blood at 7 days after PCR diagnosis of SARS-CoV-2. Test specificity was 92% for IgM and 100% for IgG. There was no significant difference in detecting IgM and IgG with Biolidics LFI and ELISA at D0 and D7 (p = 1.00), except for detection of IgM at D7 (p = 0.04). Capillary blood of SARS-CoV-2 patients showed 93% sensitivity for antibody detection. ConclusionsClinical performance of Biolidics 2019-nCoV IgG/IgM Detection Kit is comparable to ELISA and was consistent across sample types. This provides an opportunity for decentralized rapid testing and may allow point-of-care and longitudinal self-testing for the presence of anti-SARS-CoV-2 antibodies.

Development and comparison of two nanomaterial label-based lateral flow immunoassays for the detection of five antibacterial synergists

Label is a significant factor when analyzing the performance of lateral flow immunoassays (LFIAs). Thus, this study developed two nanomaterial label-based LFIA and compared their analytical performance in practical applications.

Improved performance of lateral flow immunoassays for alpha-fetoprotein and vanillin by using silica shell-stabilized gold nanoparticles.

The sensitivity of lateral flow assays (LFA) was increased 30-fold by making use of spherical core-shell gold-silica nanoparticles (AuNP@SiO2 NPs). They can be prepared by silylation of surfactant stabilized AuNPs. The AuNP@SiO2 NPs are highly stable and can be used to label antibodies at virtually any concentration. The detection limit of an LFA for alpha-fetoprotein (AFP) can be decreased from 10ng·mL-1 to 300pg·mL-1 which makes it comparable to an enzyme-linked immunosorbent assay. To demonstrate the applicability to an immunoassay, a sandwich assay was developed for vanillin by covalent modification of the AuNP@SiO2 NPs with antibody. By using the method, vanillin can be detected visually in milk powder samples in concentrations as low as 100ng·g-1. With unique optical property and great stability, this AuNP@SiO2 endows great potential in biosensing applications. Graphical abstract Controlled growth of AuNP@SiO2. The newly prepared AuNP has a negative hydration layer. This layer is further surrounded by a bilayer of CTAB through electrostatic attraction. The hydrophobic inner layer enables the access and assembling of APTES and MTTS. After the hydrolysis of siloxane, a thin layer of silica shell is formed around AuNP.

The Role of Nanoparticle Design in Determining Analytical Performance of Lateral Flow Immunoassays.

Rapid, simple, and cost-effective diagnostics are needed to improve healthcare at the point of care (POC). However, the most widely used POC diagnostic, the lateral flow immunoassay (LFA), is ∼1000-times less sensitive and has a smaller analytical range than laboratory tests, requiring a confirmatory test to establish truly negative results. Here, a rational and systematic strategy is used to design the LFA contrast label (i.e., gold nanoparticles) to improve the analytical sensitivity, analytical detection range, and antigen quantification of LFAs. Specifically, we discovered that the size (30, 60, or 100 nm) of the gold nanoparticles is a main contributor to the LFA analytical performance through both the degree of receptor interaction and the ultimate visual or thermal contrast signals. Using the optimal LFA design, we demonstrated the ability to improve the analytical sensitivity by 256-fold and expand the analytical detection range from 3 log10 to 6 log10 for diagnosing patients with inflammatory conditions by measuring C-reactive protein. This work demonstrates that, with appropriate design of the contrast label, a simple and commonly used diagnostic technology can compete with more expensive state-of-the-art laboratory tests.