MXene supported biomimetic bilayer lipid membrane biosensor for zeptomole detection of BRCA1 gene.

A biomimetic bilayer lipid membrane supported MXene based biosensor is reported for electrochemical hybridization detection of the most prevalent and potential BC biomarker BRCA1. 2D MXene nanosheet-anchored gold nanoparticle-decorated biomimetic bilayer lipid membrane (AuNP@BLM) biosensor is used for the attachment of thiolated single-stranded DNA (HS-ssDNA) targeting hybridization detection. The interaction of biomimetic bilayer lipid membrane with 2D MXene nanosheets is explored in this work for the first time. The synergistic combination of MXene and AuNP@BLM has proven to efficiently improve the detection signal to several folds. The sensor provides hybridization signals only to the complementary DNA (cDNA) sequence with a linearity range 10 zM to 1 µM and LOD of 1 zM without the need of any further amplification. The specificity of the biosensor is validated using non-complementary (ncDNA) and double base mis-match oligonucleotide DNA (dmmDNA) sequences. The sensor successfully distinguishes the signal for different target DNAs with good reproducibility indicated by the RSD value of 4.9%. Hence, we envision that the reported biosensor can be used to construct efficient diagnostic point-of-care tools based on molecular affinity interactions.

Electrochemical label free sensing of human IgG - Protein A interaction.

A novel and rapid Electrochemical Immunosensing platform was developed for the direct sensing of antibody human immuno globulin gamma (IgG) interaction with virulence factor of S. aureus, staphylococcal protein A (SpA) in the presence of electroactive redox couple ferri/ferro cyanide (K3/K4[Fe(CN)6]). The receptor SpA was attached to BioPE-DOTAP binary lipid bilayer tethered on alkane thiol molecular cushions. Atomic force microscopy (AFM), High-resolution transmission electron microscope (TEM), Fourier-transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS) techniques were used to study the molecular interactions. The AFM images showed array like formation of BioPE-DOTAP on the monolayer surface. The IgG sensor showed a linear range from 10-21 M to 10-16 M.

Carbon dots stabilized silver-lipid nano hybrids for sensitive label free DNA detection.

Carbon dots have been extensively used for the development of fluorescent based molecular affinity sensors. However, label free DNA sensing by electrochemical method is not reported so far. Herein, we report carbon dots stabilized silver nanoparticles (CD-AgNPs) lipid nano hybrids as a sensitive and selective platform for label free electrochemical DNA sensing. The CD-AgNPs were synthesized by wet chemical method and then characterized by UV-visible, Fourier-transform Infra-red (FT-IR), dynamic light scattering (DLS) and high resolution transmission electron microscopy (HR-TEM) techniques. These CD-AgNPs were used for decorating the binary lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP) surface (named as lipid) and tethered on self-assembled monolayer of 3-mercaptopropionic acid (MPA) (MPA-lipid-CD-AgNPs). The formation of array of MPA-lipid-CD-AgNPs on Au electrode was confirmed by atomic force microscopy (AFM). Electrochemical behavior of MPA- lipid-CD-AgNPs was monitored in the presence of 1 mM potassium ferri/ferrocyanide (K3/K4 [Fe(CN)6]). The formation of layer-by-layer MPA-lipid-CD-AgNPs is indicated by increased anodic and cathodic peak (ΔEp) separation with decreased redox peak current of K3/K4 [Fe(CN)6]. Short chain DNA (30 mer oligonucleotide, representing the lung cancer) was used as a model system for label free DNA sensing. Un-hybridized (single stranded DNA), hybridized (complementary hybridized), single, double and triple base mismatched target DNA hybridized surfaces were efficiently discriminated at 1 µM target DNA concentration at the Au/MPA-lipid-CD-AgNPs electrode by change in the charge transfer resistance from impedance technique. Further, the modified electrode was successfully used to determine target DNA in a wide linear range from 10-16 to 10-11 M. The present work open doors for the utilization of CDs in molecular affinity based electrochemical sensor design and development.

Supported binary liposome vesicle-gold nanoparticle for enhanced label free DNA and protein sensing.

Supported binary liposome mixture of cationic liposome N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium propane (DOTAP) and the zwitterionic liposome 1,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine (DOPE) were tethered on thiol monolayers in the absence and presence of gold nanoparticle to enhance sensor stability and sensitivity for label free DNA and protein sensing for the first time. Cysteamine hydrochloride (Cyst), 3-Mercaptopropionic acid (MPA), 11-Mercaptoundecanoic acid (MUDA) and 11-amino-1-undecane thiol (AUT) monolayers were used as tethers on gold surfaces. Electrochemical studies in the presence of [Fe(CN)6]3-/4- indicate that the presence of both DOPE and AuNP decreases the electrostatic interaction between DOTAP and MPA layer during the formation of DOPE-DOTAP-AuNP (DDA) whereas they enhance the repulsive force on the Cyst and AUT monolayers. In the thiol monolayer supported DDA, the gelation of neutral lipid DOPE by the AuNP is disfavored which inturn promotes stability of vesicle structure. The membrane protein melittin's interaction with the DDA indicates the presence of intact vesicle by showing decreased charge transfer for the MUDA and AUT in the presence of [Fe(CN)6]3-/4-. On the contrary, the presence of the bilayer and semi circled DDA on the MPA and cysteamine layers were confirmed by the increased redox reaction. Atomic Force Microscopic (AFM) and Transmission Electron Microscopic (TEM) images support the presence of an array like semi circled DDA on the MPA and well separated DDA vesicles on the MUDA with variable sizes. Dynamic Light Scattering (DLS) and Fourier Transform Infrared spectroscopy (FTIR) suggest effective coordination between DOPE, DOTAP and AuNP. Label free DNA hybridization sensing in presence of the negatively charged [Fe(CN)6]3-/4- indicates the lowest DNA detection limit of 1×10-14M with linearity range 1×10-13 to 1×10-9M. Similarly, streptavidin sensing shows the lowest detection of 1ngml-1 with a linear range 100ng to 1µg due to the increased reactive sites and distance.