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Alkyne Functionalization of a Photoactivated Ruthenium Polypyridyl Complex for Click-Enabled Serum Albumin Interaction Studies
摘要： Studying metal-protein interactions is key for understanding the fate of metallodrugs in biological systems. When a metal complex is not emissive and too weakly bound for mass spectrometry analysis, however, it may become challenging to study such interactions. In this work a synthetic procedure was developed for the alkyne functionalization of a photolabile ruthenium polypyridyl complex, [Ru(tpy)(bpy)(Hmte)](PF6)2, where tpy = 2,2′:6′,2′′-terpyridine, bpy = 2,2′-bipyridine, and Hmte = 2-(methylthio)ethanol. In the functionalized complex [Ru(HCC-tpy)(bpy)(Hmte)](PF6)2, where HCC-tpy = 4′-ethynyl-2,2′:6′,2′′-terpyridine, the alkyne group can be used for bioorthogonal ligation to an azide-labeled fluorophore using copper-catalyzed “click” chemistry. We developed a gel-based click chemistry method to study the interaction between this ruthenium complex and bovine serum albumin (BSA). Our results demonstrate that visualization of the interaction between the metal complex and the protein is possible, even when this interaction is too weak to be studied by conventional means such as UV?vis spectroscopy or ESI mass spectrometry. In addition, the weak metal complex-protein interaction is controlled by visible light irradiation, i.e., the complex and the protein do not interact in the dark, but they do interact via weak van der Waals interactions after light activation of the complex, which triggers photosubstitution of the Hmte ligand.
关键词： ruthenium polypyridyl complex,click chemistry,bovine serum albumin,photoactivation,alkyne functionalization,metal-protein interactions
更新于2025-09-23 15:21:01
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[Methods in Molecular Biology] Microfluidic Electrophoresis Volume 1906 (Methods and Protocols) || Paper-Based Electrophoresis Microchip as a Powerful Tool for Bioanalytical Applications
摘要： This chapter describes the development of paper-based microchip electrophoresis (pME) devices for the separation of clinically relevant compounds. pME were fabricated by laser cut and thermal lamination process using polyester pouches. In addition, hand-drawn pencil electrodes were integrated to the device to perform capacitively coupled contactless conductivity detection (C4D). Finished device costs less than US$ 0.10 and did not require either sophisticated instrumentation or clean room facilities. Furthermore, pME is lightweight, easy to handle, ?exible, and robust. pME-C4D device revealed an excellent capacity to separate BSA and creatinine in less than 150 s with baseline resolution. The device proposed in this chapter has proven to be a good alternative as a platform for the diagnosis of diseases from renal disorders such as diabetes mellitus and heart disease.
关键词： Biomolecules,Paper electrophoresis,Kidney failure,Pencil electrodes,Bovine serum albumin,Creatinine,Clinical diagnosis,Contactless conductivity detection
更新于2025-09-23 15:21:01
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[ACM Press the 2017 2nd International Conference - Wuhan, China (2017.11.07-2017.11.09)] Proceedings of the 2017 2nd International Conference on Communication and Information Systems - ICCIS 2017 - Effect of Concentration on Surface Acoustic Wave Velocity in A Rayleigh-type Biosensor
摘要： To predict the propagation velocity of different concentrations of bovine serum albumin (BSA) in a Rayleigh-type biosensor, we derived a new calculation method for a Rayleigh surface acoustic wave (SAW) device. The method can calculate propagation velocities of leaky Rayleigh waves and Rayleigh waves when liquid is placed in the groove of a Rayleigh-type SAW device. Different concentration of BSA proteins were used to evaluate the effect of these concentrations. In this study, we used different concentrations of BSA and a Rayleigh-type SAW device consisted of grooves of 0.25 mm width and 0.26 mm height and grooves of 0.40 mm width and 0.25 mm height, quartz as the substrate and interdigital transducers (IDT) with a 25.00 mm distance. The frequency of the Rayleigh-type SAW device was set to 157.6 MHz. Our data suggests that the propagation velocity of SAWs depends mainly upon BSA protein concentration and the size of groove. At the same concentration of BSA, the propagation acoustic wave velocity of BSA depends on the size of groove. The results showed the 0.40 mm width-0.25 mm height groove was more suitable than the 0.25 mm width-0.26 mm height groove for studying the propagation velocity of wave with liquid in a Rayleigh-type SAW device. With the propagation velocity decreased when the concentration of BSA protein increased.
关键词： Surface acoustic wave (SAW),Rayleigh-type SAW device,Leaky Rayleigh wave,Bovine serum albumin (BSA),Rayleigh wave
更新于2025-09-23 15:21:01
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Development and Validation of a Surface Plasmon Resonance Biosensor for Specific Detection of Porcine Serum Albumin in Food
摘要： Food allergies are a potential food safety and public health concern worldwide. To assure the safety of people who experience allergic reactions, it is necessary to establish effective and reliable methods for rapid detection of food allergens. This paper reports an innovative method for the rapid detection and analysis of porcine serum albumin (PSA), known as a major allergen in pork, based on a surface plasmon resonance (SPR) biosensor. The antibodies known to have a high bioactivity against PSA were verified by competitive indirect-ELISA and then immobilized on the SPR sensor surface, thus allowing them to capture PSA. The developed SPR demonstrated a linear range from 1.0 to 450 ng/mL for the measurement of PSA with a detection limit of 19.81 ng/mL. Within-day RSD (1.97–4.02%) and between-day RSD (1.88–4.15%) were no more than 5%. The SPR was evaluated for analysis of six commercial food samples and showed almost perfect agreement between the results obtained by ELISA test kits without significant differences (P > 0.05). Therefore, this assay permits accurate, specific, and sensitive detection of PSA in pork and pork products.
关键词： food safety,detection,SPR biosensor,food allergies,porcine serum albumin
更新于2025-09-23 15:21:01
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Water Soluble Fluorescent Graphene Nanodots
摘要： Water soluble fluorescent graphene nanodots (GND) have been successfully prepared through a bottom up approach from brominated pyrene via alkaline hydrothermal condensation. FT-IR, UV-Vis, Raman, XRD, AFM, and TEM studies have shown the formation of (cid:2)OH group rich nano-dots having graphene features with an approximate size of 90 nm. Steady state and 3D fluorescence studies have shown efficient fluorescence emission with maximum at 540 nm upon excitation at 450 nm in pH (cid:3) 7. Plasmid deoxyribonucleic acid (plasmid DNA) and bovine serum albumin (BSA) have shown their interaction with GND, which was studied via fluorescence quenching of GND and BSA, respectively. Selective quenching of the Fo¨ rster resonance energy transfer (FRET) of BSA-GND conjugate via ascorbic acid (AA) has been used to construct a calibration curve for AA estimation with a linear range and limit of detection of 34.00–112.00 mmole/L respectively. GND has also shown and 31.343 mmole/L, turn offchemosensing towards concentration-dependent heavy metals like Pb2 +, Cu2 +, Hg2 + and Zn2 +. In-vitromulti-color fluorescence imaging has been observed in GND-labelled HeLa (human cervical cancer) and NIH-3T3 (mouse embryonic fibroblast) cell lines with better uptake by HeLa cells. GND also has shown very high viability for both the cell lines up to the tested concentration of 300 mg/mL.
关键词： fluorescence,graphene nanodots,F?rster resonance energy transfer,brominated pyrene,bovine serum albumin
更新于2025-09-23 15:21:01
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Studies on the Interaction of Cefepime Hydrochloride with Bovine Serum Albumin by Fluorescence, Synchronous Fluorescence, Three-Dimensional Fluorescence and Circular Dichroism
摘要： The investigation on the binding mode between drug and protein is extremely important to understand biopharmaceutics, pharmacokinetics and toxicity of the drug as well as the relationship of structure and function of the protein. It is well proved that biological activity is a function of the chemical structure or structural properties. There is a study on the interaction of cefepime hydrochloride with serum albumin using in-silico molecular docking. But up to date, there is hardly any interaction investigation of cefepime hydrochloride with serum albumin utilizing by fluorescence, synchronous fluorescence, three-dimensional fluorescence and circular dichroism. In this study, the interaction of cefepime hydrochloride with bovine serum albumin in aqueous solution has been investigated by molecular spectroscopy under different pH conditions. The quenching rate constant and binding constant calculated at pH 7.4 indicated the static quenching mechanism and medium binding force. The effect of cefepime hydrochloride on the conformation of bovine serum albumin was analyzed using fluorescence, synchronous fluorescence, three-dimensional fluorescence and circular dichroism. In addition, influence of pH on the binding of cefepime hydrochloride to bovine serum albumin was investigated and the binding ability of the drug to bovine serum albumin deceased under other pH conditions (pH 1.9, 3.5, and 9.0) as compared with that at pH 7.4. As compared with the binding ability of cefepime hydrochloride to native bovine serum albumin that of cefepime hydrochloride to denatured bovine serum albumin deceases dramatically. Furthermore, the effect of metal ions on the binding constant of cefepime hydrochloride with bovine serum albumin was investigated.
关键词： Circular dichroism,Fluorescence quenching,Bovine serum albumin,Three-dimensional fluorescence,Cefepime hydrochloride
更新于2025-09-23 15:21:01
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Protein Content Sensor based on Plastic Optical Fiber
摘要： Plastic optical fiber (POF) content protein sensor has been developed. This sensor is made with a gamma and spiral configuration, with a cladding peel length of 5 cm. POF sensor is dipped into the container containing Bovine Serum Albumin (BSA) solution with different concentrations of 0.1-1 mg/ml. Both ends of the sensor are connected by Light Emiting Diode (LED) and Phototransistor. The LED light will propagate through the POF and it is received by the phototransistor. Light that propagates will be interfenced due to increasing of protein concentration around the sensor, so that the output voltage will be affected. The output voltage will be forwarded to Arduino Uno and read by the computer. The best results were obtained in a spiral configuration with 3 bending with a sensitivity of 0.007 Vml/mg and a resolution of 0.142 mg/ml. This method has advantages that are easy fabrication, simple measurement process, and low cost.
关键词： protein sensor,Plastic optical fiber,Phototransistor,Arduino Uno,Bovine Serum Albumin,LED
更新于2025-09-23 15:19:57
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Smart Assembled Human Serum Albumin Nanocarrier Enhanced Breast Cancer Treatment and Antitumor Immunity by Chemo- photothermal Therapy
摘要： High invasion and metastasis are the major obstacles to successful breast cancer therapy. Indocyanine green (ICG), a photosensitizer for photothermal therapy (PTT), shows potent anticancer efficacy when combined with the chemotherapeutic drug doxorubicin (DOX). Human serum albumin (HSA), a biocompatible carrier material, has been successfully used for the delivery of paclitaxel (Abraxane). In addition, there are ICG functional binding regions in HSA. Thus, a smart assembled nanoplatform (DI@HSA NPs) was constructed to achieve the synergistic effects of chemo-photothermal therapy against breast cancer. Compared to free ICG and free DOX, DI@HSA NPs showed satisfactory stability and exhibited an enhanced tumor targeting capacity. The mild hyperthermia generated by DI@HSA NPs can not only cause tumor photothermal ablation and promote the uptake of DI@HSA NPs by 4T1 cells, but also protect the healthy tissues nearby the tumor from overheating injury. More importantly, DI@HSA NPs greatly amplified the infiltration of CD4+ T cells and CD8+ T cells, resulting in inhibited tumor growth and metastasis. DI@HSA NPs, as a simple biocompatible nanoagent, showed excellent inhibition of breast cancer growth and metastasis by chemo-photothermal therapy, providing a potential strategy for the future therapy of breast cancer.
关键词： chemo-photothermal therapy,human serum albumin,tumor metastasis,antitumor immunity
更新于2025-09-23 15:19:57
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Ultrasensitive detection of nonlabelled bovine serum albumin using photothermal optical phase shift detection with UV excitation
摘要： Ultrasensitive detection of nonlabelled bovine serum albumin is performed in micro/nano?uidic chips using a photothermal optical phase shift (POPS) detection system. Currently, micro- and nano?uidics allow the analysis of various single cells, and their targets of interest are shifting from nucleic acids to proteins. Previously, our group developed photothermal detection techniques for the sensitive detection of non?uorescent molecules. For example, we developed a thermal lens microscope (TLM) with ultrahigh sensitivity at the single-molecule level and a POPS detector that is applicable to nanochannels smaller than the wavelength of light. The POPS detector also realized the detection of nonlabelled proteins in nanochannels, although its detection sensitivity is less than that of the TLM in microchannels due to insu?cient background light reduction. To overcome this problem, we developed a new POPS detector using relay optics for further reduction of the background light. In addition, heat transfer from the sample solution to the nanochannel wall was thoroughly investigated to achieve ultrahigh sensitivity. The limit of detection (LOD) obtained with the new POPS detector is 30 molecules in 1.0 fL. Considering this LOD, the performance of the new POPS detector is comparable with that of the TLM. Owing to the applicability of the POPS detector for sensitive detection even in nanochannels or single-μm channels, which cannot be realized with the TLM, combinations of the POPS detector and separation techniques employing unique nanochannel properties will contribute to advances in single-cell proteomics in the future.
关键词： Ultrasensitive detection,micro/nano?uidic chips,single-cell proteomics,photothermal optical phase shift,bovine serum albumin
更新于2025-09-23 15:19:57
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Rationally assembled albumin/indocyanine green nanocomplex for enhanced tumor imaging to guide photothermal therapy
摘要： Herein, a novel phototheranostic nanocomplex that is self-assembled from bovine serum albumin (BSA) and indocyanine green (ICG) is developed for enhanced near-infrared (NIR) fluorescence imaging, which benefits the guidance on in vivo cancer photothermal therapy (PTT). The study confirms that the binding of ICG with the bind sits on the albumin will result in improved hydrolytic stability and high photoluminescence quantum yield (PLQY). The ICG loading ratio in the nanocomplex is optimized and confirms the loading ratio of 0.5% ICG to be the optimal content. The optimized ICG–BSA nanocomplex (ICG–BSA NC) possesses a higher PLQY of 16.8% than that of free ICG (2.7%). The high PLQY and efficient passive targeting ability of ICG–BSA NC help improve its in vivo tumor accumulation and NIR fluorescence imaging significantly. Under laser irradiation, efficient PTT with obvious tumor growth suppression on a triple negative breast tumor model can be observed in the ICG–BSA NC treated group.
关键词： Photothermal therapy,Theranostics,Near-infrared,Indocyanine green,Albumin
更新于2025-09-23 15:19:57