Paper-based immunosensor with signal amplification by enzyme-labeled anti-p16INK4a multifunctionalized gold nanoparticles for cervical cancer screening

The aim of this study was to develop a paper-based immunosensor for cervical cancer screening, with signal amplification by multifunctionalized gold nanoparticles (AuNPs). The AuNPs were functionalized with a highly specific antibody to the p16^INK4a cancer biomarker. The signal was amplified using a combination of the peroxidase activity of horseradish peroxidase (HRP) enzyme-antibody conjugate and the peroxidase-like activity of the AuNPs. The immune complex of p16^INK4a protein and multifunctionalized AuNPs was deposited on the nitrocellulose membrane, and a positive result was generated by catalytic oxidation of peroxidase enzyme substrate 3,3’,5,5’-Tetramethylbenzidine (TMB). The entire reaction occurred on the membrane within 30 min. Evaluation in clinical samples revealed 85.2% accuracy with a kappa coefficient of 0.69. This proof of concept study demonstrates the successful development of a highly accurate, paper-based immunosensor that is easy to interpret using the naked eye and that is suitable for cervical cancer screening in low-resource settings.     

Fabrication of biocompatible magneto-fluorescence nanoparticles as a platform for fluorescent sensor and magnetic hyperthermia applications

Multifunctional nanoparticles with special magnetic and optical properties have been attracting a great deal of attention due to their important applications in the bioanalytical and biomedical fields. In this study, we report the fabrication of biocompatible magneto-fluorescence nanoparticles consisting of carbon dots (CDots) and silica-coated cobalt–manganese nanoferrites (Co_0.5Mn_0.5Fe₂O₄) (CoMnF@Si@CDots) (MagSiCDots) by a facile hydrothermal method. The as-prepared MagSiCDots have a particle size of 100–120 nm and show a negative zeta potential of −35.50 mV at a neutral pH. The fluorescence spectrum of the MagSiCDots nanoparticles consists of sharp excitation at 365 nm and broad blue light emission with a maximum wavelength of 442.5 nm and the MagSiCDots exhibit superparamagnetic behaviour with a saturation magnetization of 11.6 emu g⁻¹. The potential of MagSiCDots as a fluorescent sensor and be used for magnetic hyperthermia applications. It is seen that the fluorescent intensity of a colloidal solution (a hydrogen sulfide (H₂S) solution containing MagSiCDots nanoparticles) has a linear relationship with the H₂S concentration range of 0.2–2 μM. The limit of detection (LOD) of H₂S by our MagSiCDots particles is 0.26 μM and they remain stable for at least 90 min. To test the suitability of the MagSiCDots nanoparticles for use in hyperthermia application, induction heating using an AMF was done. It was observed that these nanoparticles had a specific absorption rate (SAR) of 28.25 W g⁻¹. The in vitro and in vivo cytotoxicity of MagSiCDots were tested on HeLa cells lines. The results show a cell viability of about 85% when exposed to 100 μg mL⁻¹ concentration of the particles. The in vivo cytotoxicity using zebrafish assay also confirmed the non-toxicity and biocompatibility of the nanoparticles to living cells. The reported data demonstrate that by combining CoMnF@Si and fluorescent CDots into a single system, not only nontoxic multifunctional nanomaterials but also multimodal nanoparticles for several applications, such as hazard gas detection and acting as a biocompatible heat source for therapeutic treatment of cancer, are provided.

Multifunctional nanoparticles with special magnetic and optical properties have been attracting a great deal of attention due to their important applications in the bioanalytical and biomedical fields.     

Pre-clinically evaluated visual lateral flow platform using influenza A and B nucleoprotein as a model and its potential applications

A visual colorimetric rapid screening system based on a lateral flow device for simultaneous detection and differentiation between influenza A and B nucleoprotein as a model was developed. Monoclonal antibodies, specific for either influenza A or B nucleoproteins, were evaluated for their reactivities and were used as targeting ligands. With the best antibody pairs selected, the system exhibited good specificity to both viruses without cross reactivity to other closely related respiratory viruses. Further semi-quantitative analysis using a strip reader revealed that the system is capable of detecting influenza A and B protein content as low as 0.04 and 1 ng per test, respectively, using a sample volume as low as 100 μL, within 10 minutes (R² = 0.9652 and 0.9718). With a performance comparison to the commercial tests, the system demonstrated a four-to-eight-fold higher sensitivity. Pre-clinical evaluation with 101 nasopharyngeal swabs reveals correlated results with a standard molecular approach, with 89% and 83% sensitivity towards influenza A and B viruses, and 100% specificity for both viruses.

Visual colorimetric rapid screening system based on lateral flow device for influenza A and B virus detection as a model and its pre-clinical evaluation.     

Hyperthermia evaluation and drug/protein-controlled release using alternating magnetic field stimuli-responsive Mn–Zn ferrite composite particles

Drug delivery particles in which the release of biomolecules is triggered by a magnetic simulant have attracted much attention and may have great potential in the fields of cancer therapy and tissue regenerative medicine. In this study, we have prepared magnetic Mn–Zn ferrite ((Mn,Zn)Fe₂O₄) (MZF) nanoparticles coated with chitosan-g-N-isopropylacrylamide (Chi-g-NIPAAm) polymer (MZF@Chi-g-NIPAAm) to deliver the anticancer drug (Doxorubicin, DOX) and bioactive proteins (Bone morphogenic protein (BMP-2)-immobilized bovine serum albumin (BSA)) (P//MZF@Chi-g-NIPAAm) and be used as chemo-hyperthermia and vector delivering biomolecules. For these purposes, we first show that the as-prepared MZF@Chi-g-NIPAAm particles exhibit super paramagnetic behavior and under certain conditions, they can act as a heat source with a specific absorption rate (SAR) of 34.88 W g⁻¹. Under acidic conditions and in the presence of AMF, the fast release of DOX was seen at around 58.9% within 20 min. In vitro evaluations indicated that concurrent thermo-chemotherapy treatment by DOX-MZF@Chi-g-NIPAAm using AMF had a better antitumor effect, compared with those using either DOX or DOX-MZF@Chi-g-NIPAAm without AMF (89.02% of cells were killed as compared to 71.82% without AMF exposure). Up to 28.18% of the BSA (used as the model protein to determine the controlled release) is released from the P//MZF@Chi-g-NIPAAm particles under AMF exposure for 1 h (only 17.31% was released without AMF). These results indicated that MZF@Chi-g-NIPAAm particles could be used to achieve hyperthermia at a precise location, effectively enhancing the chemotherapy treatments, and have a promising future as drug or bioactive delivering molecules for cancer treatment and cartilage or bone regenerative applications.

Drug delivery particles in which the release of biomolecules is triggered by a magnetic simulant have attracted much attention and may have great potential in the fields of cancer therapy and tissue regenerative medicine.