Highly Sensitive Two-Dimensional Paper Network Incorporating Biotin–Streptavidin for the Detection of Malaria

Abstract

Recently, two-dimensional paper networks have been developed to enable multistep assays to be performed in a lateral flow format. These devices have been used to perform simple enzyme linked immunoassays on paper. However, these devices have yet to incorporate more complex immunoassays, including the use of streptavidin-biotin detection strategies. Here we present a modified two-dimensional paper network capable of consecutively delivering six reagents. The device requires only a single user step and delivers (i) the sample, (ii) the biotinylated detection antibody, (iii) streptavidin horse-radish peroxidase (iv) a wash buffer (v) a colorimetric substrate and (vi) a final wash buffer. To demonstrate the utility of this approach we designed an assay to detect the malaria protein Pf HRP2. Using this platform, we were able to achieve a limit-of-detection equivalent to that of a traditional 96-well plate sandwich ELISA. In addition to improvements in the limit-of-detection, the inclusion of streptavidin-biotin simplifies the development of similar tests for other targets.

Sensitivity enhancement in lateral flow assays: a systems perspective

“…For example, improving the enzyme amplification reaction by forming the detection reagent from a biotin-antibody to streptavidin-poly-HRP reaction at the test line allowed a Pf HRP2 LFA to show equivalent sensitivity to a commercial ELISA due to the fast, strong, and multivalent biotin-streptavidin interaction. 84 The use of multivalent poly-HRP also contributed to the signal amplification. Likewise, the use of a viral nanoparticle as a multivalent detection reagent scaffold, to which multiple HRPs and multiple analyte-specific antibodies were conjugated, improved sensitivity 100-fold in a norovirus GI.1 (Norwalk) LFA 40 and nearly 1000-fold in a bacteriophage MS2 LFA.…”

Mixture of PLA–PEG and biotinylated albumin enables immobilization of avidins on electrospun fibers

“…The resulting fibrous membrane shows fast immobilization of avidin molecules, which open the door for paper-based biosensors. [38][39][40] Recently, avidin enabling release from biotinylated materials has been described, offering a potential way to develop regenerable biosensors. 27,41,42 In conclusion, biotinylated BSA surface functionalized PLA-PEG blend fibers were successfully prepared by electrospinning technique.…”

Recent Advances of Fluid Manipulation Technologies in Microfluidic Paper-Based Analytical Devices (μPADs) toward Multi-Step Assays

“…The intermediate washing steps are also essential. Several studies have been conducted to implement ELISA in paper-based devices [36,55,58,[120][121][122], as shown in Table 7. Table 7.…”