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We have developed a highly efficient technique of magnetically controlled swift loading and release of doxorubicin (DOX) drug using a magnetoelectric nanogenerator (MENG). Core-shell nanostructured MENG with a magnetostrictive core and piezoelectric shell act as field-responsive nanocarriers and possess the capability of field-triggered drug release in a cancerous environment. MENGs generate a surface electric dipole when subjected to a magnetic field due to the strain-mediated magnetoelectric effect. The capability of directional magnetic field-assisted modulation of the surface electrical dipole of MENG provides a mechanism to create/break ionic bonds with DOX molecules, which facilitates efficient drug attachment and on-demand swift detachment of the drug at a targeted site. The magnetic field-assisted drug-loading mechanism was minutely analyzed using spectrophotometry and Raman spectroscopy. The detailed time-dependent analysis of controlled drug release by the MENG under unidirectional and rotating magnetic field excitation was conducted using field-emission scanning electron microscopy, energy-dispersive X-ray, and atomic force microscopic measurements. In vitro, experiments validate the cytocompatibility and magnetically assisted on-demand and swift DOX drug delivery by the MENG near MCF-7 breast cancer cells, which results in a significant enhancement of cancer cell killing efficiency. A state-of-the-art experiment was performed to visualize the nanoscale magnetoelectric effect of MENG using off-axis electron holography under Lorentz conditions.
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The formation of clusters in non-aromatic molecules can give rise to unconventional luminescence or clusteroluminescence. Typically containing heteroatoms without extended conjugation or aromatic rings, these molecules have drawn much attention owing to the prospects of label-free biological imaging. However, their applications have been limited due to the lack of knowledge of the underlying mechanism. Herein, we have elucidated the mechanism of clusteroluminescence from proteins, which were explicitly aggregated using plasmonic silver nanoparticles. The nanoparticles promoted protein aggregation and induced nitrile formation on the surface, which, along with other lone-pair-containing heteroatoms, contributed to enhanced emission in the visible range. Remarkably, this makes imaging of proteins possible with visible excitations, as co-factor-lacking proteins generally undergo electronic transitions only in the ultraviolet range. Furthermore, the inherent protein-aggregating behaviour of plasmonic nanoparticles was harnessed for imaging of intracellular Huntingtin protein aggregates overexpressed in HeLa cells through clusteroluminescence. Significant plasmon-enhanced and red-shifted fluorescence emission was observed, which helped in the imaging and localization of the intracellular aggregates. Density functional theory calculations and transient absorbance spectroscopy were used to probe the molecular interactions at the protein-nanoparticle interface and the charge transfer states, further elucidating the role of nanoparticles and the emission mechanism. This technique thus opens alternate avenues for label-free fluorescence bioimaging.
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Nanopartículas Metálicas , Prata , Humanos , Células HeLa , Nanopartículas Metálicas/química , Prata/química , Agregados Proteicos , Luminescência , Medições LuminescentesRESUMO
Mn-doped Bi3O4Br has been synthesized using a solvothermal route. The undoped Bi3O4Br and Mn-Bi3O4Br materials possess orthorhombic unit cells with two distinct Bi sites forming a layered atomic arrangement. The shift in the (020) plane in the powder X-ray diffraction (PXRD) pattern confirms Mn-doping in the Bi3O4Br lattice. Elemental mapping indicated 7% Mn doping in the Bi3O4Br lattice structure. A core-level X-ray photoelectron study (XPS) indicates the presence of BiIII and MnII valence-states in Mn-Bi3O4Br. Doping with a cation (MnII) containing a different charge and ionic radius resulted in vacancy/defects in Mn-Bi3O4Br which further altered its electronic structure by reducing the indirect band gap, beneficial for electron conduction and electrocatalysis. The irreversible MnII to MnIII transformation at a potential of 1.48 V (vs. RHE) precedes the electrochemical oxygen evolution reaction (OER). The Mn-doped electrocatalyst achieved 10 mA cm-2 current density at 337 mV overpotential, while the pristine Bi3O4Br required 385 mV overpotential to reach the same activity. The pronounced OER activity of the Mn-Bi3O4Br sample over the pristine Bi3O4Br highlights the necessity of MnII doping. The superior activity of the Mn-Bi3O4Br catalyst over that of Bi3O4Br is due to a low Tafel slope, better double-layer capacitance (Cdl), and small charge-transfer resistance (Rct). The chronoamperometry (CA) study depicts long-term stability for 12 h at 20 mA cm-2. An electrolyzer fabricated as Pt(-)/(+)Mn-Bi3O4Br can deliver 10 mA cm-2 at a cell potential of 2.05 V. The post-CA-OER analyses of the anode confirmed the leaching of [Br-] followed by in situ formation of Mn-doped Bi2O3 as the electrocatalytically active species. Herein, an ultra-low Mn-doping into Bi3O4Br leads to an improvement in the electrocatalytic performance of the inactive Bi3O4Br material.
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Protein unfolding and aggregation are often correlated with numerous diseases such as Alzheimer's, Parkinson's, Huntington's, and other debilitating neurological disorders. Such adverse events consist of a plethora of competing mechanisms, particularly interactions that control the stability and cooperativity of the process. However, it remains challenging to probe the molecular mechanism of protein dynamics such as aggregation, and monitor them in real-time under physiological conditions. Recently, Raman spectroscopy and its plasmon-enhanced counterparts, such as surface-enhanced Raman spectroscopy (SERS) and tip-enhanced Raman spectroscopy (TERS), have emerged as sensitive analytical tools that have the potential to perform molecular studies of functional groups and are showing significant promise in probing events related to protein aggregation. We summarize the fundamental working principles of Raman, SERS, and TERS as nondestructive, easy-to-perform, and fast tools for probing protein dynamics and aggregation. Finally, we highlight the utility of these techniques for the analysis of vibrational spectra of aggregation of proteins from various sources such as tissues, pathogens, food, biopharmaceuticals, and lastly, biological fouling to retrieve precise chemical information, which can be potentially translated to practical applications and point-of-care (PoC) devices. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Diagnostic Tools > Diagnostic Nanodevices Nanotechnology Approaches to Biology > Nanoscale Systems in Biology.
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Nanotecnologia , Análise Espectral Raman , Análise Espectral Raman/métodos , Nanotecnologia/métodosRESUMO
Global proteome changes in microbes affect the survival and overall production of commercially relevant metabolites through different bioprocesses. The existing methods to monitor proteome level changes are destructive in nature. Stable isotope probing (SIP) coupled with Raman spectroscopy is a relatively new approach for proteome analysis. However, applying this approach for monitoring changes in a large culture volume is not cost-effective. In this study, for the first time we are presenting a novel method of combining reverse SIP using 13 C-glucose and Deuterium to monitor the proteome changes through Raman spectroscopy. The findings of the study revealed visible changes (blue shifts) in proteome related peaks that can be used for monitoring proteome dynamics, that is, synthesis of nascent amino acids and its turnover with time in a non-destructive, cost-effective, and label-free manner.
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Proteoma , Análise Espectral Raman , Proteoma/metabolismo , Análise Espectral Raman/métodos , Marcação por Isótopo/métodos , Proteômica , Escherichia coliRESUMO
The quest to enhance Raman spectroscopic signals through the rational design of plasmonic substrates has enabled the detection and characterization of pharmaceutically important molecules with low scattering cross-sections, such as amino acids and proteins, and is helping in making forays into the diverse field of biomedical sciences. This work presents a simple strategy for synthesizing silver nanoparticles-incorporated alumina nanofibers (Ag-AlNFs) utilizing controlled microwave synthesis for enhancing the surface-enhanced Raman chemical enhancement factor through photo-induced charge accumulation at the plasmonic-dielectric interface. The plasmonic-dielectric fibers serve as excellent charge carrier trappers, as evident from the ultrafast transient absorption spectroscopy studies. Apart from chemical enhancement, the increase in electronic surface charge also enables the protein disulfide bonds to capture these electrons and form a transient disulfide electron adduct radical, which converts to free thiol radical on dissociation. This allows protein molecules to bind to the nanoparticle's surface with the favorable silver thiol bond leading to greater surface affinity and larger SERS enhancement. The proposed Ag-AlNFs represent a cost-effective material that can be potentially used to probe biological systems in a label-free manner by photoactivating the SERS substrate for obtaining higher enhancement factors.
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Deep-fat frying of food develops lipid oxidation products that deteriorate oil and pose a health risk. This necessitates the development of a rapid and accurate oil quality and safety detection technique. Herein, surface-enhanced Raman spectroscopy (SERS) and sophisticated chemometric techniques were used for rapid and label-free determination of peroxide value (PV) and fatty acid composition of oil in-situ. In the study, plasmon-tuned and biocompatible Ag@Au core-shell nanoparticle-based SERS substrates were used to obtain optimum enhancement despite matrix interference to efficiently detect the oil components. The potent combination of SERS and the Artificial Neural Network (ANN) method could determine the fatty acid profile and PV with upto 99% accuracy. Moreover, the SERS-ANN method could quantify the low level of trans fats, i.e., < 2%, with 97% accuracy. Therefore, the developed algorithm-assisted SERS system enabled the sleek and rapid monitoring and on-site detection of oil oxidation.
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Nanopartículas Metálicas , Nanopartículas , Análise Espectral Raman/métodos , Ácidos Graxos , Oxirredução , Algoritmos , Nanopartículas Metálicas/químicaRESUMO
Although medical advances have increased our grasp of the amazing morphological, genetic, and phenotypic diversity of diseases, there are still significant technological barriers to understanding their complex and dynamic character. Specifically, the complexities of the biological systems throw a diverse set of challenges in developing efficient theranostic tools and methodologies that can probe and treat pathologies. Among several emerging theranostic techniques such as photodynamic therapy, photothermal therapy, magnetic resonance imaging, and computed tomography, Raman spectroscopy (RS) is emerging as a promising tool that is a label-free, cost-effective, and non-destructive technique. It can also provide real-time diagnostic information and can employ multimodal probes for detection and therapy. These attributes make it a perfect candidate for the analytical counterpart of the existing theranostic probes. The use of biocompatible nanomaterials for the fabrication of Raman probes provides rich structural information about the biological molecules, cells, and tissues and highly sensitive information down to single-molecule levels when integrated with advanced RS tools. This review discusses the fundamentals of Raman spectroscopic tools such as surface-enhanced Raman spectroscopy and Resonance Raman spectroscopy, their variants, and the associated theranostic applications. Besides the advantages, the current limitations, and future challenges of using RS in disease diagnosis and therapy have also been discussed.
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Nanoestruturas , Fotoquimioterapia , Medicina de Precisão , Análise Espectral Raman , Nanoestruturas/uso terapêutico , NanotecnologiaRESUMO
Surface plasmon resonance (SPR) has the ability to drive catalytic conversion of the reactant molecules via the production of hot electrons, which in general requires high activation energy. The reactions driven by these hot electrons are critical and essential in various heterogeneous surface catalytic reactions. However, there is a need to understand the dynamics of surface reactions and the underlying mechanism, which are influenced by several factors such as the constitution of the nanoparticle, exposure time, and reaction conditions to name a few. However, the effect of solvent in stabilizing the electron-hole pair, the orientation, and the surface coverage of the analyte are poorly understood due to the limitations of current methods. To get deeper insights into the reaction dynamics, we have demonstrated the combined utility of plasmon-enhanced Raman spectroscopy and Two-dimensional correlation spectroscopy (2DCOS) to study the plasmon-driven conversion of 4-nitrothiophenol on the surface of plasmonic nanoparticles. Interestingly, this combined technique provided us with previously unobservable results regarding surface catalysis by conventional spectroscopic analysis alone. Specifically, for the first time, 2DCOS provided critical insights in bridging the gap in our understanding of the interplay of solvent effect, orientation, and surface packing of the analyte molecules. It was observed that certain species like 4,4-dimercaptoazobenzene (DMAB) or 4-aminothiophenol (4-ATP) can be selectively formed based on the ordered or disordered phases of the analytes on the surface, thus paving the way to precisely control light-driven reactions through operando spectroscopy.
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The emergence of nanoparticles in biomedical applications has made their interactions with proteins inevitable. Nanoparticles conjugated with proteins and peptide-based constructs form an integral part of nanotherapeutics and have recently shown promise in treating a myriad of diseases. The proper functioning of proteins is critical to achieve their biological functions. However, interface issues result in the denaturation of proteins, and the loss of orientation and steric hindrance can adversely affect the function of the conjugate. Furthermore, surface-induced denaturation also triggers protein aggregation, resulting in amyloid-like species. Understanding the mechanistic underpinnings of protein-nanoparticle interactions and controlling their interfacial characteristics are critical and challenging due to the complex nature of the conjugates. In this milieu, we demonstrate that ionic liquids can be suitable candidates for stabilizing protein-nanoparticle interactions by virtue of their excellent protein-preserving properties. We also probe the previously unexplored mechanism of ion-mediated stabilization of the protein molecules on the nanoparticle surface. The protein-nanoparticle conjugates consist of lysozyme and choline-based ionic liquids characterized by optical and electron microscopy techniques combined with surface-sensitive plasmon-enhanced Raman spectroscopy. Furthermore, atomistic molecular dynamics simulations of the conjugates delineate interfacial interactions of the protein molecules and the modulation by the ions, particularly the conformational changes and the dynamic correlation when the protein and specific ionic liquid molecules are adsorbed on the nanoparticle surface. The combined experimental and computational studies showed the synergistic behavior of the ions of the ionic liquids, specifically the orientation and coverage of the anions aided by the cations to control the surface interactions and hence the overall protein stability. These studies pave the way for using ionic liquids, particularly their biocompatible counterparts in nanoparticle-based complexes, as stabilizing agents for biomedical applications.
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Abnormal protein kinetics could be a cause of several diseases associated with essential life processes. An accurate understanding of protein dynamics and turnover is essential for developing diagnostic or therapeutic tools to monitor these changes. Raman spectroscopy in combination with stable isotope probes (SIP) such as carbon-13, and deuterium has been a breakthrough in the qualitative and quantitative study of various metabolites. In this work, we are reporting the utility of Raman-SIP for monitoring dynamic changes in the proteome at the community level. We have used 13 C-labeled glucose as the only carbon source in the medium and verified its incorporation in the microbial biomass in a time-dependent manner. A visible redshift in the Raman spectral vibrations of major biomolecules such as nucleic acids, phenylalanine, tyrosine, amide I, and amide III were observed. Temporal changes in the intensity of these bands demonstrating the feasibility of protein turnover monitoring were also verified. Kanamycin, a protein synthesis inhibitor was used to assess the feasibility of identifying effects on protein turnover in the cells. Successful application of this work can provide an alternate/adjunct tool for monitoring proteome-level changes in an objective and nondestructive manner.
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Carbono , Proteoma , Análise Espectral Raman/métodos , Glucose/metabolismoRESUMO
Fluorescence imaging is a major driver of discovery in biology, and an invaluable asset in clinical diagnostics. To overcome quenching limitations of conventional fluorescent dyes and further improve intensity, nanoparticle-based constructs have been the subject of intense investigation, and within this realm, dye-doped silica-coated nanoparticles have garnered significant attention. Despite their growing popularity in research, fluorescent silica nanoparticles suffer from a significant flaw. The degradation of these nanoparticles in biological media by hydrolytic dissolution is underreported, leading to serious misinterpretations, and limiting their applicability for live cell and in vivo imaging. Here, the development of an ultra-stable, dye-embedded, silica-coated metal nanoparticle is reported, and its superior performance in long-term live cell imaging is demonstrated. While conventional dye-doped silica nanoparticles begin to degrade within an hour in aqueous media, by leveraging a modified liquid calcination process, this new construct is shown to be stable for at least 24 h. The stability of this metal-enhanced fluorescent probe in biologically relevant temperatures and media, and its demonstrated utility for cell imaging, paves the way for its future adoption in biomedical research.
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Nanopartículas Metálicas , Nanopartículas , Fluorescência , Corantes Fluorescentes , Dióxido de Silício , ÁguaRESUMO
A generalized label-free platform for surface-selective molecular sensing in living cells can transform the ability to examine complex events in the cell membrane. While vertically aligned semiconductor and metal-semiconductor hybrid nanopillars have rapidly surfaced for stimulating and probing the intracellular environment, the potential of such constructs for selectively interrogating the cell membrane is surprisingly underappreciated. In this work, a new platform, entitled nano-PROD (nano-pillar based Raman optical detection), enables molecular recording by probing fundamental vibrational modes of membrane constituents of cells adherent on a large-area silver-coated silicon nanopillar substrate fabricated using a precursor solution-based nanomanufacturing process. It is shown that the nano-PROD platform sustains live cells in near-physiological conditions, which can be directly profiled using surface-enhanced Raman spectroscopy due to the confined electromagnetic field enhancement. The experimental results highlight the utility of the platform in probing specific cell surface markers for accurately recognizing the phenotypically identical prostate cancer cells, differing only in prostate-specific membrane antigen expression. Due to its tunability, nano-PROD has the promise to be readily extendable to other applications that can leverage its unique combination of nanoscale topographic features and molecular sensing capabilities, from stain-free cytopathology inspection to understanding spatio-mechanical regulation in membrane receptor function.
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Prata , Análise Espectral Raman , Membrana Celular , SilícioRESUMO
We demonstrate the remarkable ability of surface-enhanced Raman spectroscopy (SERS) to track the allosteric changes in restriction endonuclease KpnI (R.KpnI) caused by metal ions. R.KpnI binds and promiscuously cleaves DNA upon activation by Mg2+ ions. However, the divalent ion Ca2+ induces high fidelity cleavage, which can be overcome by higher concentrations of Mg2+ ions. In the absence of any 3D crystal structure, for the first time, we have elucidated the structural underpinnings of such a differential effect of divalent ions on the endonuclease activity. A combined SERS and molecular dynamics (MD) approach showed that Ca2+ ion activates an enzymatic switch in the active site, which is responsible for the high fidelity activity of the enzyme. Thus, SERS in combination with MD simulations provides a powerful tool for probing the link between the structure and activity of enzyme molecules that play vital roles in DNA transactions.
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Clivagem do DNA , Análise Espectral Raman , Cátions Bivalentes , Desoxirribonucleases de Sítio Específico do Tipo II/genética , ÍonsRESUMO
Olsalazine (Olsa) is a broad-spectrum anti-cancer agent acting as a DNA-methylation inhibitor. When conjugated to 2-cyano-6-aminobenzothiazole and a peptide substrate specific for the tumor-overexpressed enzyme furin, it can self-assemble into nanoparticles that can be detected by chemical-exchange saturation-transfer magnetic-resonance imaging (CEST MRI). We report here that these nano-assemblies can also be detected with high specificity in furin-overexpressing tumor cells by Raman spectroscopy with a distinct scattering signature and demonstrate the utility of this sensing mechanism inâ vitro and inâ vivo. Our findings suggest that Raman spectroscopy could be used for high-resolution image-guided surgery to precisely delineate tumor margins during and after resection in real-time as well as to determine microscopic tumor invasion and multifocal locoregional tumor spread, which are currently impossible to visualize with available imaging technologies, including CEST MRI.
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Ácidos Aminossalicílicos/química , Imageamento por Ressonância Magnética/métodos , Nanopartículas/química , Neoplasias/diagnóstico por imagem , Animais , Meios de Contraste/química , Células HCT116 , Humanos , Camundongos , Camundongos SCID , Microscopia de Fluorescência , Neoplasias/patologia , Análise Espectral Raman , Transplante HeterólogoRESUMO
The use of surface-enhanced Raman spectroscopy (SERS) to determine spectral markers for the diagnosis of heparin-induced thrombocytopenia (HIT), a difficult-to-diagnose immune-related complication that often leads to limb ischemia and thromboembolism, is proposed. The ability to produce distinct molecular signatures without the addition of labels enables unbiased inquiry and makes SERS an attractive complementary diagnostic tool. A capillary-tube-derived SERS platform offers ultrasensitive, label-free measurement as well as efficient handling of blood serum samples. This shows excellent reproducibility, long-term stability and provides an alternative diagnostic rubric for the determination of HIT by leveraging machine-learning-based classification of the spectroscopic data. We envision that a portable Raman instrument could be combined with the capillary-tube-based SERS analytical tool for diagnosis of HIT in the clinical laboratory, without perturbing the existing diagnostic workflow.
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Heparina/efeitos adversos , Análise Espectral Raman , Trombocitopenia/induzido quimicamente , Trombocitopenia/diagnóstico , Humanos , Aprendizado de Máquina , Fatores de TempoRESUMO
Among the strategies used for enhancement of tumour retention of imaging agents or anticancer drugs is the rational design of probes that undergo a tumour-specific enzymatic reaction preventing them from being pumped out of the cell. Here, the anticancer agent olsalazine (Olsa) was conjugated to the cell-penetrating peptide RVRR. Taking advantage of a biologically compatible condensation reaction, single Olsa-RVRR molecules were self-assembled into large intracellular nanoparticles by the tumour-associated enzyme furin. Both Olsa-RVRR and Olsa nanoparticles were readily detected with chemical exchange saturation transfer magnetic resonance imaging by virtue of exchangeable Olsa hydroxyl protons. In vivo studies using HCT116 and LoVo murine xenografts showed that the OlsaCEST signal and anti-tumour therapeutic effect were 6.5- and 5.2-fold increased, respectively, compared to Olsa without RVRR, with an excellent 'theranostic correlation' (R2 = 0.97) between the imaging signal and therapeutic response (normalized tumour size). This furin-targeted, magnetic resonance imaging-detectable platform has potential for imaging tumour aggressiveness, drug accumulation and therapeutic response.
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Ácidos Aminossalicílicos/metabolismo , Antineoplásicos/metabolismo , Furina/metabolismo , Espaço Intracelular/metabolismo , Imageamento por Ressonância Magnética/métodos , Nanopartículas , Ácidos Aminossalicílicos/química , Animais , Antineoplásicos/química , Catálise , Linhagem Celular Tumoral , Transformação Celular Neoplásica , Células HCT116 , Humanos , CamundongosRESUMO
Nanoparticle-based drug delivery systems have attracted significant interest owing to their promise as tunable platforms that offer improved intracellular release of cargo therapeutics. However, significant challenges remain in maintaining the physiological stability of the mucosal matrix due to the nanoparticle-induced reduction in the matrix diffusivity and promotion of mucin aggregation. Such aggregation also adversely impacts the permeability of the nanoparticles, and thus, diminishes the efficacy of nanoparticle-based formulations. Here, an entirely complementary approach is proposed to the existing nanoparticle functionalization methods to address these challenges by using trehalose, a naturally occurring disaccharide that offers exceptional protein stabilization. Plasmon-enhanced Raman spectroscopy and far-red fluorescence emission of the plasmonic silver nanoparticulate clusters are harnessed to create a unique dual-functional, aggregating, and imaging agent that obviates the need of an additional reporter to investigate mucus-nanoparticle interactions. These spectroscopy-based density mapping tools uncover the mechanism of mucus-nanoparticle interactions and establish the protective role of trehalose microenvironment in minimizing the nanoparticle aggregation. Thus, in contrast to the prevailing belief, these results demonstrate that nonfunctionalized nanoparticles may rapidly penetrate through mucus barriers, and by leveraging the bioprotectant attributes of trehalose, an in vivo milieu for efficient mucosal drug delivery can be generated.
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Nanopartículas Metálicas/química , Muco/metabolismo , Análise Espectral Raman/métodos , Trealose/farmacocinética , Animais , Sistemas de Liberação de Medicamentos , Humanos , Jejuno/metabolismo , Prata/química , SuínosRESUMO
Imaging tip growth in fungal hyphae is highly warranted to unravel the molecular mechanism of this extraordinarily precise and localized phenomenon. Inâ situ probing of fungal cultures, however, have been challenging due to their inherent complexity and light penetration issues associated with conventional optical imaging. In this work, we report a label-free approach using a combination of light sheet microscopy and Raman spectroscopy to obtain concomitant morphological and biochemical information from the growing specimen. We show that the variance in morphology in the symbiotic fungus Piriformospora indica are rooted in the underlying differences in chemical composition in the specific growth zones. Our findings suggest that this potent two-pronged approach can comprehensively characterize growth areas and elucidate microbe interactions in still developing colonies with high sensitivity and multiplexing capability.
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Basidiomycota/química , Hifas/crescimento & desenvolvimento , Análise Espectral Raman , MicroscopiaRESUMO
Surface enhanced Raman spectroscopy (SERS) has emerged as a promising technique for the rapid and ultrasensitive detection of molecular species such as drugs of abuse in biofluids. Yet, it remains a significant challenge to create a viable screening tool for multiple drug classes, owing to the lack of affinity of certain species for the SERS substrate and to the matrix interference in complex media. Here we report a protein tethering SERS approach, which blends plasmonic enhancement with facile drug binding, to engineer a rapid, label-free and versatile screening tool for narcotics. By exploiting the known binding attributes of human serum albumin, we determine the effective concentration of narcotics present in solution through differential enhancement of the spectral markers. In conjunction with chemometric methods, this approach not only enables unambiguous recognition of different drug classes, such as barbiturates, opiates, amphetamines and benzodiazepines, but also offers a lower limit of detection in comparison to direct SERS application. Through molecular docking simulations, we probe the mechanistic underpinnings of the protein tethering approach paving the way for narcotic detection in clinical samples in the near future.