scholarly journals Rapid and Easy-to-Use Method for Accurate Characterization of Target Binding and Kinetics of Magnetic Particle Bioconjugates for Biosensing

Sensors ◽  
2021 ◽  
Vol 21 (8) ◽  
pp. 2802
Author(s):  
Averyan V. Pushkarev ◽  
Alexey V. Orlov ◽  
Sergey L. Znoyko ◽  
Vera A. Bragina ◽  
Petr I. Nikitin
Keyword(s):  
Optical Sensors ◽  
Magnetic Particle ◽  
Troponin I ◽  
Sample Volume ◽  
Label Free ◽  
Spectral Correlation ◽  
Faster Development ◽  
Selection Of

The ever-increasing use of magnetic particle bioconjugates (MPB) in biosensors calls for methods of comprehensive characterization of their interaction with targets. Label-free optical sensors commonly used for studying inter-molecular interactions have limited potential for MPB because of their large size and multi-component non-transparent structure. We present an easy-to-use method that requires only three 20-min express measurements to determine the key parameters for selection of optimal MPB for a biosensor: kinetic and equilibrium characteristics, and a fraction of biomolecules on the MPB surface that are capable of active targeting. The method also provides a prognostic dependence of MPB targeting efficiency upon interaction duration and sample volume. These features are possible due to joining a magnetic lateral flow assay, a highly sensitive sensor for MPB detection by the magnetic particle quantification technique, and a novel mathematical model that explicitly describes the MPB-target interactions and does not comprise parameters to be fitted additionally. The method was demonstrated by experiments on MPB targeting of cardiac troponin I and staphylococcal enterotoxin B. The validation by an independent label-free technique of spectral-correlation interferometry showed good correlation between the results obtained by both methods. The presented method can be applied to other targets for faster development and selection of MPB for affinity sensors, analytical technologies, and realization of novel concepts of MPB-based biosensing in vivo.

scholarly journals Preclinical Testing of Radiopharmaceuticals for the Detection and Characterization of Osteomyelitis: Experiences from a Porcine Model

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4221
Author(s):  
Aage Kristian Olsen Alstrup ◽  
Svend Borup Jensen ◽  
Ole Lerberg Nielsen ◽  
Lars Jødal ◽  
Pia Afzelius
Keyword(s):  
Animal Model ◽  
Animal Models ◽  
Porcine Model ◽  
Animal Experiments ◽  
Radioactive Tracers ◽  
The Individual ◽  
Selection Of

The development of new and better radioactive tracers capable of detecting and characterizing osteomyelitis is an ongoing process, mainly because available tracers lack selectivity towards osteomyelitis. An integrated part of developing new tracers is the performance of in vivo tests using appropriate animal models. The available animal models for osteomyelitis are also far from ideal. Therefore, developing improved animal osteomyelitis models is as important as developing new radioactive tracers. We recently published a review on radioactive tracers. In this review, we only present and discuss osteomyelitis models. Three ethical aspects (3R) are essential when exposing experimental animals to infections. Thus, we should perform experiments in vitro rather than in vivo (Replacement), use as few animals as possible (Reduction), and impose as little pain on the animal as possible (Refinement). The gain for humans should by far exceed the disadvantages for the individual experimental animal. To this end, the translational value of animal experiments is crucial. We therefore need a robust and well-characterized animal model to evaluate new osteomyelitis tracers to be sure that unpredicted variation in the animal model does not lead to a misinterpretation of the tracer behavior. In this review, we focus on how the development of radioactive tracers relies heavily on the selection of a reliable animal model, and we base the discussions on our own experience with a porcine model.

Characterization of Tissue Microstructure Scatterer Distribution with Spectral Correlation

1993 ◽  
Vol 15 (3) ◽  
pp. 238-254 ◽  
Author(s):  
Tomy Varghese ◽  
Kevin D. Donohue
Keyword(s):  
Formation Process ◽  
Biological Tissue ◽  
Statistical Parameters ◽  
Spectral Correlation ◽  
Tissue Microstructure ◽  
Power Spectral ◽  
Spectral Peaks ◽  
Ultrasound Signal

Characterization of tissue microstructure from the backscattered ultrasound signal using the spectral autocorrelation (SAC) function provides information about the scatterer distribution in biological tissue. This paper demonstrates SAC capabilities in characterizing periodicities in A-scans due to regularity in the scatterer distribution. The A-scan is modelled as a cyclostationary signal, where the statistical parameters of the signal vary in time with single or multiple periodicities. This periodicity manifests itself as spectral peaks both in the power spectral density (PSD) and in the SAC. Periodicity in the PSD will produce a well defined dominant peak in the cepstrum, which has been used to determine the scatterer spacing. The relationship between the scatterer spacing and the spacing of the spectral peaks is established using a stochastic model of the echo-formation process from biological tissue. The distribution of the scatterers within the microstructure is modelled using a Gamma function, which offers a flexible method of simulating parametric regularity in the scatterer spacing. Simulations of the tissue microstructure for lower orders of regularity indicate that the SAC components reveal information about the scatterer spacing that are not seen in the PSD and the cepstrum. The echo-formation process is tested by simulating microstructure of varying regularity and analyzing their effect on the SAC, PSD and cepstrum. Experimental validation of the simulation results are provided using in vivo scans of the breast and liver tissue that show the presence of significant spectral correlation components in the SAC.

In vivo selection of phage sequences and characterization of peptide-specific binding to breast cancer cells

2008 ◽  
Vol 5 (2) ◽  
pp. 128-131 ◽  
Author(s):  
Rui Wang ◽  
Lin Zhang ◽  
Hongkai Zhang ◽  
Xiyin Wei ◽  
Yi Yang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Specific Binding ◽  
In Vivo Selection ◽  
Selection Of

Characterization of two in vivo challenge models to measure functional activity of monoclonal antibodies to Plasmodium falciparum circumsporozoite protein

2020 ◽  
Author(s):  
Rama Raghunandan ◽  
Bryan T Mayer ◽  
Yevel Flores-Garcia ◽  
Monica W Gerber ◽  
Raphael Gottardo ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Monoclonal Antibodies ◽  
Functional Activity ◽  
Statistical Power ◽  
Rank Order ◽  
Circumsporozoite Protein ◽  
Dose Selection ◽  
Selection Of

Abstract Background New strategies are needed to reduce the incidence of malaria, and promising approaches include the development of vaccines and monoclonal antibodies (mAbs) that target the circumsporozoite protein (CSP). To select the best candidates and speed development, it is essential to standardize preclinical assays to measure the potency of such interventions in animal models. Methods Two assay configurations were studied using transgenic Plasmodium berghei expressing Plasmodium falciparum full-length circumsporozoite protein. The assays measured 1) reduction in parasite infection of the liver (liver burden) following an intravenous (i.v) administration of sporozoites and 2) protection from parasitaemia following mosquito bite challenge. Two human CSP mAbs, AB311 and AB317, were compared for their ability to inhibit infection. Multiple independent experiments were conducted to define assay variability and resultant impact on the ability to discriminate differences in mAb functional activity. Results Overall, the assays produced highly consistent results in that all individual experiments showed greater functional activity for AB317 compared to AB311 as calculated by the dose required for 50% inhibition (ID50) as well as the serum concentration required for 50% inhibition (IC50). The data were then used to model experimental designs with adequate statistical power to rigorously screen, compare, and rank order novel anti-CSP mAbs. Conclusion The results indicate that in vivo assays described here can provide reliable information for comparing the functional activity of mAbs. The results also provide guidance regarding selection of the appropriate experimental design, dose selection, and group sizes.

Epidemiological and molecular characterization ofStaphylococcus haemolyticusstrains, from a hematology ward, with decreased susceptibility to glycopeptides

2011 ◽  
Vol 57 (6) ◽  
pp. 476-484 ◽  
Author(s):  
Xiao Xue Ma ◽  
Dan Dan Sun ◽  
Jian Hu ◽  
En Hua Wang ◽  
En Jie Luo
Keyword(s):  
Minimum Inhibitory Concentration ◽  
Molecular Characterization ◽  
Continuous Monitoring ◽  
Inhibitory Concentration ◽  
Pulsed Field Gel Electrophoresis ◽  
Gene Complex ◽  
Case Patient ◽  
Selection Of

In the present study, we report on the reduced susceptibility to teicoplanin among clinical isolates of Staphylococcus haemolyticus in a hematology ward of a teaching hospital. The molecular characterization of 17 S. haemolyticus strains was performed using mec gene complex classification, pulsed-field gel electrophoresis analysis, and minimum inhibitory concentration examination. Pulsotype A strains carrying a class C2 mec gene complex were the most prevalent strains, at 64.7%. In vivo selection of stepwise increase in resistance to vancomycin and teicoplanin was observed in three S. haemolyticus strains serially isolated from a case patient. The results of the present study suggest the regional spread of certain S. haemolyticus clones with diminished susceptibility to glycopeptides, emphasizing the need for continuous monitoring of minimum inhibitory concentration levels of vancomycin and teicoplanin in S. haemolyticus strains, and the importance of infection control practices to prevent its transmission.

scholarly journals Label-free visualization and characterization of extracellular vesicles in breast cancer

2019 ◽  
Vol 116 (48) ◽  
pp. 24012-24018 ◽  
Author(s):  
Sixian You ◽  
Ronit Barkalifa ◽  
Eric J. Chaney ◽  
Haohua Tu ◽  
Jaena Park ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Progression ◽  
Metabolic Imaging ◽  
Live Cells ◽  
Clinical Samples ◽  
Label Free ◽  
Human Breast

Despite extensive interest, extracellular vesicle (EV) research remains technically challenging. One of the unexplored gaps in EV research has been the inability to characterize the spatially and functionally heterogeneous populations of EVs based on their metabolic profile. In this paper, we utilize the intrinsic optical metabolic and structural contrast of EVs and demonstrate in vivo/in situ characterization of EVs in a variety of unprocessed (pre)clinical samples. With a pixel-level segmentation mask provided by the deep neural network, individual EVs can be analyzed in terms of their optical signature in the context of their spatial distribution. Quantitative analysis of living tumor-bearing animals and fresh excised human breast tissue revealed abundance of NAD(P)H-rich EVs within the tumor, near the tumor boundary, and around vessel structures. Furthermore, the percentage of NAD(P)H-rich EVs is highly correlated with human breast cancer diagnosis, which emphasizes the important role of metabolic imaging for EV characterization as well as its potential for clinical applications. In addition to the characterization of EV properties, we also demonstrate label-free monitoring of EV dynamics (uptake, release, and movement) in live cells and animals. The in situ metabolic profiling capacity of the proposed method together with the finding of increasing NAD(P)H-rich EV subpopulations in breast cancer have the potential for empowering applications in basic science and enhancing our understanding of the active metabolic roles that EVs play in cancer progression.

scholarly journals Toward Development of a Label-Free Detection Technique for Microfluidic Fluorometric Peptide-Based Biosensor Systems

Micromachines ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 691
Author(s):  
Nikita Sitkov ◽  
Tatiana Zimina ◽  
Alexander Kolobov ◽  
Vladimir Karasev ◽  
Alexander Romanov ◽  
...  
Keyword(s):  
Troponin I ◽  
Troponin T ◽  
Detection System ◽  
Label Free ◽  
Protein Markers ◽  
Peptide Aptamers ◽  
Registration System ◽  
Biosensor System ◽  
Label Free Detection ◽  
Selection Of

The problems of chronic or noncommunicable diseases (NCD) that now kill around 40 million people each year require multiparametric combinatorial diagnostics for the selection of effective treatment tactics. This could be implemented using the biosensor principle based on peptide aptamers for spatial recognition of corresponding protein markers of diseases in biological fluids. In this paper, a low-cost label-free principle of biomarker detection using a biosensor system based on fluorometric registration of the target proteins bound to peptide aptamers was investigated. The main detection principle considered includes the re-emission of the natural fluorescence of selectively bound protein markers into a longer-wavelength radiation easily detectable by common charge-coupled devices (CCD) using a specific luminophore. Implementation of this type of detection system demands the reduction of all types of stray light and background fluorescence of construction materials and aptamers. The latter was achieved by careful selection of materials and design of peptide aptamers with substituted aromatic amino acid residues and considering troponin T, troponin I, and bovine serum albumin as an example. The peptide aptamers for troponin T were designed in silico using the «Protein 3D» (SPB ETU, St. Petersburg, Russia) software. The luminophore was selected from the line of ZnS-based solid-state compounds. The test microfluidic system was arranged as a flow through a massive of four working chambers for immobilization of peptide aptamers, coupled with the optical detection system, based on thick film technology. The planar optical setup of the biosensor registration system was arranged as an excitation-emission cascade including 280 nm ultraviolet (UV) light-emitting diode (LED), polypropylene (PP) UV transparent film, proteins layer, glass filter, luminophore layer, and CCD sensor. A laboratory sample has been created.

scholarly journals Biochemical Profiles of In Vivo Oral Mucosa by Using a Portable Raman Spectroscopy System

Optics ◽  
2021 ◽  
Vol 2 (3) ◽  
pp. 134-147
Author(s):  
Marcelo Saito Nogueira ◽  
Victoria Ribeiro ◽  
Marianna Pires ◽  
Felipe Peralta ◽  
Luis Felipe das Chagas e Silva de Carvalho
Keyword(s):  
Raman Spectroscopy ◽  
Biochemical Characterization ◽  
Principal Component ◽  
Histopathological Analysis ◽  
Label Free ◽  
Healthy Individuals ◽  
Normal Tissues ◽  
The Difference

Most oral injuries are diagnosed by histopathological analysis of invasive and time-consuming biopsies. This analysis and conventional clinical observation cannot identify biochemically altered tissues predisposed to malignancy if no microstructural changes are detectable. With this in mind, detailed biochemical characterization of normal tissues and their differentiation features on healthy individuals is important in order to recognize biomolecular changes associated with early tissue predisposition to malignant transformation. Raman spectroscopy is a label-free method for characterization of tissue structure and specific composition. In this study, we used Raman spectroscopy to characterize the biochemistry of in vivo oral tissues of healthy individuals. We investigated this biochemistry based on the vibrational modes related to Raman spectra of four oral subsites (buccal, gingiva, lip and tongue) of ten volunteers as well as with principal component (PC) loadings for the difference between the four types of oral subsites. Therefore, we determined the biochemical characteristics of each type of healthy oral subsite and those corresponding to differentiation of the four types of subsites. In addition, we developed a spectral reference of oral healthy tissues of individuals in the Brazilian population for future diagnosis of early pathological conditions using real-time, noninvasive and label-free techniques such as Raman spectroscopy.

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