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1.
The current study presents a new miniature microfluidic flow cytometer integrated with several functional micro-devices capable of viral sample purification and detection by utilizing a magnetic bead-based immunoassay. The magnetic beads were conjugated with specific antibodies, which can recognize and capture target viruses. Another dye-labeled anti-virus antibody was then used to mark the bead-bound virus for the subsequent optical detection. Several essential components were integrated onto a single chip including a sample incubation module, a micro flow cytometry module and an optical detection module. The sample incubation module consisting of pneumatic micropumps and a membrane-type, active micromixer was used for purifying and enriching the target virus-bound magnetic beads with the aid of a permanent magnet. The micro flow cytometry module and the optical detection module were used to perform the functions of virus counting and collection. Experimental results showed that virus samples with a concentration of 10(3)PFU/ml can be automatically detected successfully by the developed system. In addition, the entire diagnosis procedure including sample incubation and virus detection took only about 40min. Consequently, the proposed micro flow cytometry may provide a powerful platform for rapid diagnosis and future biological applications. 相似文献
2.
Rachel Legmann H. Brett Schreyer Rodney G. Combs Ellen L. McCormick A. Peter Russo Seth T. Rodgers 《Biotechnology and bioengineering》2009,104(6):1107-1120
Multi‐factorial experimentation is essential in understanding the link between mammalian cell culture conditions and the glycoprotein product of any biomanufacturing process. This understanding is increasingly demanded as bioprocess development is influenced by the Quality by Design paradigm. We have developed a system that allows hundreds of micro‐bioreactors to be run in parallel under controlled conditions, enabling factorial experiments of much larger scope than is possible with traditional systems. A high‐throughput analytics workflow was also developed using commercially available instruments to obtain product quality information for each cell culture condition. The micro‐bioreactor system was tested by executing a factorial experiment varying four process parameters: pH, dissolved oxygen, feed supplement rate, and reduced glutathione level. A total of 180 micro‐bioreactors were run for 2 weeks during this DOE experiment to assess this scaled down micro‐bioreactor system as a high‐throughput tool for process development. Online measurements of pH, dissolved oxygen, and optical density were complemented by offline measurements of glucose, viability, titer, and product quality. Model accuracy was assessed by regressing the micro‐bioreactor results with those obtained in conventional 3 L bioreactors. Excellent agreement was observed between the micro‐bioreactor and the bench‐top bioreactor. The micro‐bioreactor results were further analyzed to link parameter manipulations to process outcomes via leverage plots, and to examine the interactions between process parameters. The results show that feed supplement rate has a significant effect (P < 0.05) on all performance metrics with higher feed rates resulting in greater cell mass and product titer. Culture pH impacted terminal integrated viable cell concentration, titer and intact immunoglobulin G titer, with better results obtained at the lower pH set point. The results demonstrate that a micro‐scale system can be an excellent model of larger scale systems, while providing data sets broader and deeper than are available by traditional methods. Biotechnol. Bioeng. 2009; 104: 1107–1120. © 2009 Wiley Periodicals, Inc. 相似文献
3.
Micro sequential injection system as the interfacing device for process analytical applications
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It is an important and desirable capability to be able to control the quality and quantity of biological product by maintaining and adjusting bioreactor performance throughout its production duration. Amino acids are the building blocks of proteins. Scientists will need to ensure sufficient supply of amino acids as the substrates in the bioreactors as well as to control the excess level of undesirable free amino acid byproducts to maintain an optimum growth environment for cell culture. We have developed a compact and robust sample preparation platform capable of interfacing with analytical instruments to achieve bioreactor amino acids monitoring. We demonstrated the feasibility of this concept by incorporating an automatic amino acid sample preparation protocol to a micro sequential injection (μSI) system connected to an ultra‐performance liquid chromatography system for real‐time, at‐line amino acid separation, and quantitation. The μSI system was configured into a “platform‐like” sample preparation system that is able to accommodate future wet chemistry‐type sample preparations. Its real‐time amino acid results can be readily available to bioprocess scientists for quick decision making and design of their next experiment. Potential automatic feedback control mechanisms can be established through trigger events based on predetermined analytical signal thresholds so the system can communicate with facility infrastructure to control bioreactors in near real‐time fashion. The proposed μSI system described in this paper can be widely used as an automatic sample preparation system connected to the front‐end of analytical instruments to enable process analytical technology applications. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:607–613, 2015 相似文献
4.
Data Processing System (DPS) software with experimental design,statistical analysis and data mining developed for use in entomological research 总被引:27,自引:0,他引:27
Abstract A comprehensive but simple‐to‐use software package called DPS (Data Processing System) has been developed to execute a range of standard numerical analyses and operations used in experimental design, statistics and data mining. This program runs on standard Windows computers. Many of the functions are specific to entomological and other biological research and are not found in standard statistical software. This paper presents applications of DPS to experimental design, statistical analysis and data mining in entomology. 相似文献
5.
Saturnin Ombinda‐Lemboumba Rudzani Malabi Masixole Y. Lugongolo Lebogang Thobakgale Sello L. Manoto Patience Mthunzi‐Kufa 《Journal of biophotonics》2019,12(7)
Transmission measurement has been perceived as a potential candidate for label‐free investigation of biological material. It is a real‐time, label‐free and non‐invasive optical detection technique that has found wide applications in pharmaceutical industry as well as the biological and medical fields. Combining transmission measurement with optical trapping has emerged as a powerful tool allowing stable sample trapping, while also facilitating transmittance data analysis. In this study, a near‐infrared laser beam emitting at a wavelength of 1064 nm was used for both optical trapping and transmission measurement investigation of human immunodeficiency virus 1 (HIV‐1) infected and uninfected TZM‐bl cells. The measurements of the transmittance intensity of individual cells in solution were carried out using a home built optical trapping system combined with laser transmission setup using a single beam gradient trap. Transmittance spectral intensity patterns revealed significant differences between the HIV‐1 infected and uninfected cells. This result suggests that the transmittance data analysis technique used in this study has the potential to differentiate between infected and uninfected TZM‐bl cells without the use of labels. The results obtained in this study could pave a way into developing an HIV‐1 label‐free diagnostic tool with possible applications at the point of care . 相似文献
6.
Jürgen B Barken KB Tobisch S Pioch D Wümpelmann M Hecker M Schweder T 《Biotechnology and bioengineering》2005,92(3):299-307
The knowledge of critical process-relevant genes can be used for an improved control of bioprocesses. So far bioprocess-relevant marker genes can be analyzed by established expression analysis methods only off-line. In this study, an alternative approach for a potential at-line monitoring of gene expression during bioprocesses is suggested. This approach is based on the measurement of specific mRNAs on an electric DNA-chip in connection with a magnetic bead-based sandwich hybridization. In order to allow an at-line measurement of specific mRNAs an improved method for a fast and partially automated isolation of high quality-RNA samples was developed. The expression analysis of the electric DNA-chip was compared with optical DNA micro arrays and the real time RT-PCR for three selected process-relevant genes of Bacillus subtilis. We demonstrate that the mRNA analysis by means of the electric DNA-chip gives similar results compared to the micro array analysis and the real time RT-PCR technique. 相似文献
7.
In this work, we developed a motion estimation and correction method which real-time obtained the direction and displacement of repetitive micro bulk motion (such as cardiac and respiratory motion) on an SS-OCT system without additional tracking hardware, and reduced the motion noise in optical coherence tomography angiography (OCTA). In the approach, the direction of repetitive micro bulk motion was considered fixed, and proportional relationships between the motion components in three directions were determined; Then we performed one-dimension cross-correlation to obtain depth displacement which was further used to obtain other two motion components, and greatly reduced the computation; The processing speed on a graphic processing unit was 478 pairs of B-Scans per second, and the measurement range was larger than the range of the angiogram-based methods. Lastly, corrupt angiograms were recovered by adaptive scan protocol, and reduced acquisition time in comparison with the previous work. 相似文献
8.
Ford SM Davies J Kar B Qi SD McWhorter S Soper SA Malek CK 《Journal of biomechanical engineering》1999,121(1):13-21
Micromachining was performed in polymethylmethacrylate (PMMA) using X-ray lithography for the fabrication of miniaturized devices (microchips) for potential applications in chemical and genetic analyses. The devices were fabricated using two different techniques: transfer mask technology and a Kapton mask. For both processes, the channel topography was transferred (1:1) to the appropriate substrate via the use of an optical mask. In the case of the transfer mask technique, the PMMA substrate was coated with a positive photoresist and a thin Au/Cr plating base. Following UV exposure, the resist was developed and a thick overlayer (approximately 3 microns) of Au electroplated onto the PMMA substrate only where the resist was removed, which acted as an absorber of the X-rays. In the other technique, a Kapton film was used as the X-ray mask. In this case, the Kapton film was UV exposed using the optical mask to define the channel topography and following development of the resist, a thick Au overlayer (8 microns) was electrodeposited onto the Kapton sheet. The PMMA wafer during X-ray exposure was situated directly underneath the Kapton mask. In both cases, the PMMA wafer was exposed to soft X-rays and developed to remove the exposed PMMA. The resulting channels were found to be 20 microns in width (determined by optical mask) with channel depths of approximately 50 microns (determined by x-ray exposure time). In order to demonstrate the utility of this micromachining process, several components were fabricated in PMMA including capillary/chip connectors, injectors for fixed-volume sample introduction, separation channels for electrophoresis and integrated fiber optic fluorescence detectors. These components could be integrated into a single device to assemble a system appropriate for the rapid analysis of various targets. 相似文献
9.
A Streptococcus uberis transposon mutant screen reveals a negative role for LiaR homologue in biofilm formation
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T. Salomäki T. Karonen P. Siljamäki K. Savijoki T.A. Nyman P. Varmanen A. Iivanainen 《Journal of applied microbiology》2015,118(1):1-10
Deinococcus spp are among the most radiation‐resistant micro‐organisms that have been discovered. They show remarkable resistance to a range of damage caused by ionizing radiation, desiccation, UV radiation and oxidizing agents. Traditionally, Escherichia coli and Saccharomyces cerevisiae have been the two platforms of choice for engineering micro‐organisms for biotechnological applications, because they are well understood and easy to work with. However, in recent years, researchers have begun using Deinococcus spp in biotechnologies and bioremediation due to their specific ability to grow and express novel engineered functions. More recently, the sequencing of several Deinococcus spp and comparative genomic analysis have provided new insight into the potential of this genus. Features such as the accumulation of genes encoding cell cleaning systems that eliminate organic and inorganic cell toxic components are widespread among Deinococcus spp. Other features such as the ability to degrade and metabolize sugars and polymeric sugars make Deinococcus spp. an attractive alternative for use in industrial biotechnology. 相似文献
10.
Impact of media and antifoam selection on monoclonal antibody production and quality using a high throughput micro‐bioreactor system
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Sai Rashmika Velugula‐Yellela Abasha Williams Nicholas Trunfio Chih‐Jung Hsu Brittany Chavez Seongkyu Yoon Cyrus Agarabi 《Biotechnology progress》2018,34(1):262-270
Monoclonal antibody production in commercial scale cell culture bioprocessing requires a thorough understanding of the engineering process and components used throughout manufacturing. It is important to identify high impact components early on during the lifecycle of a biotechnology‐derived product. While cell culture media selection is of obvious importance to the health and productivity of mammalian bioreactor operations, other components such as antifoam selection can also play an important role in bioreactor cell culture. Silicone polymer‐based antifoams were known to have negative impacts on cell health, production, and downstream filtration and purification operations. High throughput screening in micro‐scale bioreactors provides an efficient strategy to identify initial operating parameters. Here, we utilized a micro‐scale parallel bioreactor system to study an IgG1 producing CHO cell line, to screen Dynamis, ProCHO5, PowerCHO2, EX‐Cell Advanced, and OptiCHO media, and 204, C, EX‐Cell, SE‐15, and Y‐30 antifoams and their impacts on IgG1 production, cell growth, aggregation, and process control. This study found ProCHO5, EX‐Cell Advanced, and PowerCHO2 media supported strong cellular growth profiles, with an IVCD of 25‐35 × 106 cells‐d/mL, while maintaining specific antibody production (Qp > 2 pg/cell‐d) for our model cell line and a monomer percentage above 94%. Antifoams C, EX‐Cell, and SE‐15 were capable of providing adequate control of foaming while antifoam 204 and Y‐30 noticeably stunted cellular growth. This work highlights the utility of high throughput micro bioreactors and the importance of identifying both positive and negative impacts of media and antifoam selection on a model IgG1 producing CHO cell line. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:262–270, 2018 相似文献
11.
Kimimasa Tobita Xiaoqin Liu Cecilia W. Lo 《Birth defects research. Part C, Embryo today : reviews》2010,90(3):176-184
Assessment of structural birth defects (SBDs) in animal models usually entails conducting detailed necropsy for anatomical defects followed by histological analysis for tissue defects. Recent advances in new imaging technologies have provided the means for rapid phenotyping of SBDs, such as using ultra‐high frequency ultrasound biomicroscopy, optical coherence tomography, micro‐CT, and micro‐MRI. These imaging modalities allow the detailed assessment of organ/tissue structure, and with ultrasound biomicroscopy, structure and function of the cardiovascular system also can be assessed noninvasively, allowing the longitudinal tracking of the fetus in utero. In this review, we briefly discuss the application of these state‐of‐the‐art imaging technologies for phenotyping of SBDs in rodent embryos and fetuses, showing how these imaging modalities may be used for the detection of a wide variety of SBDs. Birth Defects Research (Part C) 90:176–184, 2010. © 2010 Wiley‐Liss, Inc. 相似文献
12.
Yang Wang Peng Wang Xue Zhou Chang Li Huizeng Li Xiaotian Hu Fengyu Li Xiaoping Liu Mingzhu Li Yanlin Song 《Liver Transplantation》2018,8(12)
Achieving light harvesting is crucial for the efficiency of the solar cell. Constructing optical structures often can benefit from micro‐nanophotonic imprinting. Here, a simple and facile strategy is developed to introduce a large area grating structure into the perovskite‐active layer of a solar cell by utilizing commercial optical discs (CD‐R and DVD‐R) and achieve high photovoltaic performance. The constructed diffraction grating on the perovskite active layer realizes nanophotonic light trapping by diffraction and effectively suppresses carrier recombination. Compared to the pristine perovskite solar cells (PSCs), the diffraction‐grating perovskite devices with DVD obtain higher power conversion efficiency and photocurrent density, which are improved from 16.71% and 21.67 mA cm?2 to 19.71% and 23.11 mA cm?2. Moreover, the stability of the PSCs with diffraction‐grating‐structured perovskite active layer is greatly enhanced. The method can boost photonics merge into the remarkable perovskite materials for various applications. 相似文献
13.
Aims: To determine the ability of a bismuth thiol to control floc formation in a multispecies population of micro‐organisms obtained from the activated sludge unit of a wastewater treatment plant. The molecular level mechanisms by which bismuth‐2‐3‐dimercapto‐1‐propanol (BisBAL) inhibits bioaggregation are also elucidated. Methods and Results: Micro‐organisms were grown over a 3‐day period in a batch system by adding glucose as an electron donor to stimulate short‐term heterotrophic activity. Extracellular polymeric substances (EPS) produced by activated sludge micro‐organisms during exponential and stationary growth phases in the presence and absence of BisBAL were characterized using colorimetry, X‐ray photoelectron spectroscopy (XPS), and Fourier transform infrared (FTIR) spectroscopy. BisBAL at its minimum inhibitory concentration (MIC, 10 μmol l?1) was most effective in suppressing microbial floc formation. The principal effect of sub‐inhibitory concentrations of BisBAL was to decrease total EPS production while largely preserving homology. Conclusions: Antifouling and bactericidal properties of BisBAL arise from its ability to reduce EPS expression and preferentially suppressing acidic and O‐acetylated carbohydrates and certain protein secondary structures viz. β‐structures, random coils, and α‐and 3‐turn helices. As micro‐organisms exhibited a much weaker tendency to aggregate at lower concentrations of these specific EPS components, they also appear to be important for the formation of microbial flocs and bioaggregates. Significance and Impact of the Study: BisBAL was shown to be highly effective against multispecies microbial aggregation. Novel bismuth‐based biocides could also be potentially employed to control excess sludge production in wastewater treatment systems by inhibiting EPS expression. 相似文献
14.
Status and future of disease protection and grape berry quality alteration by micro‐organisms in viticulture
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Grapevine is one of the most widely grown fruit crops in the world. At present, however, there is much concern regarding chemical pollution in viticulture due to the application of chemical fungicides and fertilizers. One viticultural practice to resolve this issue is the application of micro‐organisms to grapevine as a substitute for chemicals. Some micro‐organisms act as an enhancer of grape berry quality as well as a suppresser of disease in grapevine through their antagonistic ability and/or systemic resistance inducing ability. Herein, we review current and prospective applications of micro‐organisms in viticulture.
Significance and Impact of the Study
In this review, we evaluate the applicability of micro‐organisms in viticulture. Micro‐organisms can improve grape berry quality through grapevine disease protection and grape berry quality alteration. Because the use of micro‐organisms to protect grapevine from plant diseases is safer than the use of chemical fungicides, the use of biofungicides in viticulture is expected to be enhanced by the increasing consumer concern towards chemical fungicides. Micro‐organisms also modify plant secondary metabolites for use as flavours, pharmaceuticals and food additives. Studies of micro‐organisms that promote polyphenol, anthocyanin and aroma compound biosynthesis are in progress with an eye to improving grape berry quality. 相似文献15.
Boyi Chen Jun Peng Heping Shen The Duong Daniel Walter Steve Johnston Mowafak M. Al‐Jassim Klaus J. Weber Thomas P. White Kylie R. Catchpole Daniel Macdonald Hieu T. Nguyen 《Liver Transplantation》2019,9(7)
A fast, nondestructive, camera‐based method to capture optical bandgap images of perovskite solar cells (PSCs) with micrometer‐scale spatial resolution is developed. This imaging technique utilizes well‐defined and relatively symmetrical band‐to‐band luminescence spectra emitted from perovskite materials, whose spectral peak locations coincide with absorption thresholds and thus represent their optical bandgaps. The technique is employed to capture relative variations in optical bandgaps across various PSCs, and to resolve optical bandgap inhomogeneity within the same device due to material degradation and impurities. Degradation and impurities are found to both cause optical bandgap shifts inside the materials. The results are confirmed with micro‐photoluminescence spectroscopy scans. The excellent agreement between the two techniques opens opportunities for this imaging concept to become a quantified, high spatial resolution, large‐area characterization tool of PSCs. This development continues to strengthen the high value of luminescence imaging for the research and development of this photovoltaic technology. 相似文献
16.
Amy K. Manocchi Peter Domachuk Fiorenzo G. Omenetto Hyunmin Yi 《Biotechnology and bioengineering》2009,103(4):725-732
The rapid development in optical detection techniques for sensing applications has led to an increased need for biocompatible, biodegradable, and disposable optical components. We present a controllable fabrication technique for an entirely biopolymeric planar optical waveguide via simple spin‐coating. The refractive index difference, thermal responsive properties, and inherent biocompatibility of gelatin and agarose were exploited in the fabrication of thin, stacked films that efficiently guide light in a core layer with higher index of refraction. These planar waveguides were fabricated using a simple spin‐coating technique, which resulted in controllable layer thicknesses and smooth layer interfaces. This technique, therefore, offers a path for routine engineering of biopolymer structures with contrasting refractive indices. The thermal stability of the gelatin core layer was improved using two crosslinkers; glutaraldehyde or microbial Transglutaminase. Light guiding in the core layer of the waveguide was demonstrated using a simple He–Ne laser setup. Guiding efficiency was further illustrated by directly embedding fluorescent markers within the core layer and detecting their spectral signature. Combined with the biopolymers' inherent biocompatibility and biodegradability, our simple strategy to fabricate disposable optical components holds the potential for the development of applications in biological sensing and implantable biomedical devices. Biotechnol. Bioeng. 2009;103: 725–732. © 2009 Wiley Periodicals, Inc. 相似文献
17.
18.
One major challenge currently facing the biopharmaceutical industry is to understand how MAb microheterogeneity affects therapeutic efficacy, potency, immunogenicity, and clearance. MAb micro‐heterogeneity can result from post‐translational modifications such as sialylation, galactosylation, C‐terminal lysine cleavage, glycine amidation, and tryptophan oxidation, each of which can generate MAb charge variants; such heterogeneity can affect pharmacokinetics (PK) considerably. Implementation of appropriate on‐line quality control strategies may help to regulate bioprocesses, thus enabling more homogenous material with desired post‐translational modifications and PK behavior. However, one major restriction to implementation of quality control strategies is the availability of techniques for obtaining on‐line or at‐line measurements of these attributes. In this work, we describe the development of an at‐line assay to separate MAb charge variants in near real‐time, which could ultimately be used to implement on‐line quality control strategies for MAb production. The assay consists of a 2D‐HPLC method with sequential in‐line Protein A and WCX‐10 HPLC column steps. To perform the 2D‐HPLC assay at‐line, the two columns steps were integrated into a single method using a novel system configuration that allowed parallel flow over column 1 or column 2 or sequential flow from column 1 to column 2. A bioreactor system was also developed such that media samples could be removed automatically from bioreactor vessels during production and delivered to the 2D‐HPLC for analysis. With this at‐line HPLC assay, we have demonstrated that MAb microheterogeneity occurs throughout the cell cycle whether the host cell line is grown under different or the same nominal culture conditions. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:249–255, 2014 相似文献
19.
Pobozy E Radomska A Koncki R Głab S 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2003,789(2):417-424
Micellar electrokinetic chromatography with UV absorbance detection has been applied for fast and selective determination of creatinine in samples of postdialysate fluid. Optimization of the method was performed, with the best results being obtained using a 30 mM borate-100 mM sodium dodecyl sulphate background electrolyte, pH 9, with the detector set at 235 nm and an applied voltage of 17 kV across a fused-silica capillary of 67 cm/75 micro m I.D. The linear range of the technique was over 2 orders of magnitude (5-1000 micro M). The developed analytical procedure is useful for the monitoring of clinical hemodialysis treatment, because creatinine levels in real undiluted samples of postdialysate range from 80 to 350 micro M. The separation system allows the analysis of about six to seven samples of spent dialysate per hour in almost real time. The determinations are not influenced by other components of dialysate fluid nor by other surrogates extracted from patient blood. The results of analysis using the developed procedure and the kinetic spectrophotometric Jaffe method conventionally used in clinical settings for creatinine determination are fully comparable. Successful clinical evaluation of the analytical system was performed. The developed system is useful for bloodless estimation of bioanalytical parameters of hemodialysis sessions such as creatinine-time profiles and total creatinine removal. Both these parameters are important in clinical models of hemodialysis therapy. 相似文献
20.
Liwang Liu Laurent Plawinski Marie‐Christine Durrieu Bertrand Audoin 《Journal of biophotonics》2019,12(8)
Advances in microscopy with new visualization possibilities often bring dramatic progress to our understanding of the intriguing cellular machinery. Picosecond optoacoustic micro‐spectroscopy is an optical technique based on ultrafast pump‐probe generation and detection of hypersound on time durations of picoseconds and length scales of nanometers. It is experiencing a renaissance as a versatile imaging tool for cell biology research after a plethora of applications in solid‐state physics. In this emerging context, this work reports on a dual‐probe architecture to carry out real‐time parallel detection of the hypersound propagation inside a cell that is cultured on a metallic substrate, and of the hypersound reflection at the metal/cell adhesion interface. Using this optoacoustic modality, several biophysical properties of the cell can be measured in a noncontact and label‐free manner. Its abilities are demonstrated with the multiple imaging of a mitotic macrophage‐like cell in a single run experiment. 相似文献