Improving the Sensitivity of Nanofibrous Membrane-Based ELISA for On-Site Antibiotics Detection

Abstract

An ultrasensitive and portable colorimetric enzyme-linked immunosorbent assay (ELISA) sensor for antibiotics was fabricated by immobilizing antibodies inside the largely porous and highly hydrophilic nanofibrous membranes. Different from regular electrospun nanofibrous membranes where antibodies may frequently be blocked by the heterogeneous porous structure and sterically crowded loaded on the surface, the controlled microporous structure and increased hydrophilicity of nanofibrous membranes could improve the diffusion properties of antibodies, reduce the sterically crowding effect, and dramatically improve the sensitivity of the membrane-based ELISA. The limitation of detection (LOD) for chloramphenicol (CAP) reached 0.005 ng/mL, around 200 times lower than the conventional paper-based ELISA, making quantitative analysis and portable on-site detection achievable via the use of smartphones. The successful design and fabrication of the nanofibrous membrane-based ELISA with novel features overcome the structural drawbacks of regular electrospun nanofibrous membranes and provide new paths to develop highly sensitive on-site detection of hazardous chemical agents.

Highly Sensitive Naked Eye Detectable Colorimetric Biosensors Made from Macroporous Framework Melamine Foams for Onsite and Simultaneous Detection of Multiple Environmental Hazards in Flowing Through Sensing Systems

“…The red channel (R) values from RGB values represent the color intensity. [19,53] Here, the red channel intensity change could be represented by ΔRGB value, which was obtained by the RGB value difference between the white background and each membrane, as the equation of ΔRGB = RGB background − RGB membranes (1) Statistical Analysis: All experiments were repeated three times. Data are expressed as mean ± standard deviations (SD).…”

“…[18] Such a structure is perfect for serving as filters, however, not ideal for use as biological sensor materials, since diffusion of large biomolecules through NF, such as HIgG could be significantly reduced, leading to reduced amounts of proteins loaded onto the inside surfaces of NF membranes. [19] Different from the NF membrane materials, the MF framework structure can allow diffusion and penetration of large molecules through the membranes at high speed and freedom. As demonstrated in Figure 1, there is no size exclusion effect in the MF.…”

“…[14,15,16,17] Such a structural feature could consequently lower the sensitivity of ELISA sensors made by nitrocellulose membranes, filter papers, and even nanofibrous membranes, resulting in inhomogeneous colorimetric signals of p-ELISA sensors. [18,19,20] Thus, an ideal media with a three-dimensional homogenous and open macroporous structure that can allow large biomolecules migrate freely inside in all directions was envisioned as a better material for such ELISA biosensors. Macroporous aerogels have some 3D structural features.…”

Multi-Analyte Discriminated ECL via Photonic Crystal-Enhanced Selectivity

“…Different antibiotics, including β-lactams, quinolone antibiotics, aminoglycosides, macrolides antibiotics, etc., − are widely used in livestock industry, − agricultural farmin, , medical industry, aquaculture, , and food products. , The abuse of antibiotics increases the mutation frequency of drug-resistant genes, which is seriously influencing and jeopardizing the ecological environment, medical treatment, human health, food safety, and other aspects (Figure a). High-performance liquid chromatography, , chromatography-mass spectrometry, and enzyme-linked immunosorbent assay , were adopted for the detections of antibiotics, but containing expensive apparatus, time-consuming, and complicate sample pretreatment process. As the most critical medicine for clinic treatment or animal husbandry, it calls the efficient and convenient method for the multi-analytes antibiotics detection in complex system of realistic environments or in vivo samples.…”

Radial mesoporous silica supported Eu3+‐functionalized carbon dots for ratiometric tetracycline detection and their polyvinylidene fluoride hybrid film for temperature sensing

“…3 Hence, it is of great significance to develop a handy, rapid, and accurate strategy for TC detection. So far, the applicable techniques for TC sensing mainly include enzyme-linked immunosorbent assays, 4 high-performance liquid chromatography, 5 colorimetry, 6 capillary electrophoresis, 7 and fluorescence method. 8,9 Among them, the fluorescence technology with the merits of simple instrument operation, short time-consuming, low cost, and high sensitivity has received tremendous attention.…”