Luminescence properties and Judd-Ofelt analysis of Tb3+ doped Sr2YTaO6 double perovskite phosphors for white LED applications.

A series of Tb3+-doped Sr2YTaO6 double perovskite phosphors (SYT:Tb3+) were synthesized using a conventional solid-state reaction method. A strong green emission was observed in the SYT:Tb3+ phosphors, and the optimal doping concentration of Tb3+ was confirmed to be 5 mol%. The electric dipole-dipole interaction was ascribed to be the main mechanism for the luminescence concentration quenching. Analysis of the concentration-dependent fluorescence decay confirmed that the self-generated quenching model holds for the dynamic process of Tb3+ decays in SYT. Furthermore, the internal quantum efficiencies, non-radiative transition rates, and energy transfer rates of the 5D4 level for the SYT:Tb3+ samples were estimated, respectively. The luminescence thermal stability of the sample was also evaluated based on the Arrhenius model. The chromaticity shift of the SYT:5 mol% Tb3+ phosphor was examined to be 0.013 when the sample temperature was increased from 303 to 483 K, thus indicating excellent chromaticity shifting resistance under high temperature conditions. Moreover, the Judd-Ofelt parameters were calculated from the emission spectra of SYT:Tb3+ to be Ω2 = 0.29 × 10-20, Ω4 = 0.45 × 10-20, and Ω6 = 0.72 × 10-20 cm2, respectively. The fluorescence branching ratios and radiative transition rates for the 5D4 level were calculated based on the obtained Judd-Ofelt parameters. Finally, a white light-emitting diode (LED) prototype was assembled using a 310 nm LED chip combined with a prepared green SYT:Tb3+ phosphor and two other commercial blue and red phosphors. The obtained warm white light exhibits good chromaticity coordinates (0.32, 0.32) and a high color rendering index of 96.1. Based on the above results, it can be known that the prepared SYT:Tb3+ phosphors have a potential application as green emitting phosphors in white LEDs.

Multicolor-emitting Er3+ and Er3+/Yb3+ doped Zn2GeO4 phosphors combining static and dynamic identifications for advanced anti-counterfeiting application.

Anti-counterfeiting labels based on luminescence materials are a newly emerging technique for protecting legal goods and intellectual property. In the anti-counterfeiting field to prevent forgery and cloning, luminescence materials with properties different from the commercialized and traditional ones are in urgent need. In this work, multicolor-emitting Er3+ single-doped and Er3+/Yb3+ co-doped Zn2GeO4 phosphors combining static and dynamic identifications were developed in order to achieve advanced anti-counterfeiting application. The variation of trap content with increasing the doping content of rare earth ions was analyzed through X - ray photoelectron spectroscopy, thermoluminescence analysis. It was found that there are two types of traps with different depth in Zn2GeO4 phosphors. The depths of the traps were experimentally confirmed to be 0.68 and 0.79 eV, respectively. The transient photocurrent response measurement confirmed the existence of charge carriers, and the mechanism for long persistent luminescence was deduced. The multicolor upconversion mechanisms under 980 and 1550 nm excitation were also discovered. Based on the multicolor steady and transient emission features, an anti-counterfeiting pattern was designed using the phosphors. Static and dynamic identification was demonstrated and presented in detail. Finally, it is indicated that the studied phosphors are excellent candidates for potential applications in luminescence anti-counterfeiting labels.

Near infrared emissions from both high efficient quantum cutting (173%) and nearly-pure-color upconversion in NaY(WO4)2:Er3+/Yb3+ with thermal management capability for silicon-based solar cells.

Raising photoelectric conversion efficiency and enhancing heat management are two critical concerns for silicon-based solar cells. In this work, efficient Yb3+ infrared emissions from both quantum cutting and upconversion were demonstrated by adjusting Er3+ and Yb3+ concentrations, and thermo-manage-applicable temperature sensing based on the luminescence intensity ratio of two super-low thermal quenching levels was discovered in an Er3+/Yb3+ co-doped tungstate system. The quantum cutting mechanism was clearly decrypted as a two-step energy transfer process from Er3+ to Yb3+. The two-step energy transfer efficiencies, the radiative and nonradiative transition rates of all interested 4 f levels of Er3+ in NaY(WO4)2 were confirmed in the framework of Föster-Dexter theory, Judd-Ofelt theory, and energy gap law, and based on these obtained efficiencies and rates the quantum cutting efficiency was furthermore determined to be as high as 173% in NaY(WO4)2: 5 mol% Er3+/50 mol% Yb3+ sample. Strong and nearly pure infrared upconversion emission of Yb3+ under 1550 nm excitation was achieved in Er3+/Yb3+ co-doped NaY(WO4)2 by adjusting Yb3+ doping concentrations. The Yb3+ induced infrared upconversion emission enhancement was attributed to the efficient energy transfer 4I11/2 (Er3+) + 2F7/2 (Yb3+) → 4I15/2 (Er3+) + 2F5/2 (Yb3+) and large nonradiative relaxation rate of 4I9/2. Analysis on the temperature sensing indicated that the NaY(WO4)2:Er3+/Yb3+ serves well the solar cells as thermos-managing material. Moreover, it was confirmed that the fluorescence thermal quenching of 2H11/2/4S3/2 was caused by the nonradiative relaxation of 4S3/2. All the obtained results suggest that NaY(WO4)2:Er3+/Yb3+ is an excellent material for silicon-based solar cells to improve photoelectric conversion efficiency and thermal management.

K64 acetylation of heat shock protein 90 suppresses nucleopolyhedrovirus replication in Bombyx mori.

HSP90 is a highly conserved chaperone that facilitates the proliferation of many viruses, including silkworm (bombyx mori) nucleopolyhedrovirus (BmNPV), but the underlying regulatory mechanism was unclear. We found that suppression of HSP90 by 17-AAG, a HSP90-specific inhibitor, significantly reduced the expression of BmNPV capsid protein gp64 and viral genome replication, whereas overexpression of B. mori HSP90(BmHSP90) promoted BmNPV replication. Furthermore, in a recent study of the lysine acetylome of B. mori infected with BmNPV, we focused on the reduced viral proliferation due to changes of BmHSP90 lysine acetylation. Site-directed introduction of acetylated (K/Q) or deacetylated (K/R) mimic mutations into BmHSP90 revealed that lysine 64 (K64) acetylation activated the JAK/STAT pathway and reduced BmHSP90 ATPase activity, leading to diminished chaperone activity and ultimately inhibiting BmNPV proliferation. In this study, a single lysine 64 acetylation change of BmHSP90 was elucidated as a model of posttranslational modifications occurring in the wake of host-virus interactions, providing novel insights into potential antiviral strategies.

Defect-induced Ce3+ sites preferences and multilevel concentration quenching of a high-efficiency cyan phosphor for high-quality full-visible-spectrum wLED.

Currently, the efficient way to synthesize white light-emitting diodes (WLEDs) is combining a near-ultraviolet (n-UV, 380-420 nm) emitting LED chip with tricolor (red, green, and blue) emitting phosphors. However, further improving the color rendering index (CRI) for WLEDs is hindered by the absence of cyan components. Hence, a series of high-efficiency and continuously tunable Ce3+,Gd3+-doped CaScBO4 (CSBO) blue-cyan phosphors with an orthorhombic structure were successfully developed by a high-temperature solid-state reaction method. Based on density-functional theory (DFT) calculation, a vacancy was produced along with inequivalent replacement (3Ca2+ → 2Ce3+/Gd3+ + V''Ca) when just adding the trivalent cations, meanwhile causing the local environment of the lattice to relax so Ce3+/Gd3+ ions find it easier to enter into Sc3+ sites at a higher doping concentration. Under the excitation of n-UV, the emission peak position moves from 443 nm to 480 nm and two concentration quenching points appear with an increase in Ce3+ ions by defect-induced site-selective occupation. The two samples at concentration quenching points both have high quantum efficiencies of 88.6% and 86.2% and an acceptable thermal quenching performance. The property performance and internal mechanism are illuminated by the excitation and emission spectra and theoretical analysis. Finally, by combining CSBO:Ce3+, commercial green and red phosphors and an n-UV LED chip, an as-fabricated WLED with a great CRI value of 93.2 and a low CCT (4291K) was obtained. This work demonstrates the potential of CSBO:Ce3+ as a blue-cyan phosphor for use in high-quality full-visible-spectrum WLEDs in the future. The investigation of the mechanism for the defect-induced site preferences provides a reference for developing new photoluminescent materials.

Heat shock proteins and viral infection.

Heat shock proteins (HSPs) are a kind of proteins which mostly found in bacterial, plant and animal cells, in which they are involved in the monitoring and regulation of cellular life activities. HSPs protect other proteins under environmental and cellular stress by regulating protein folding and supporting the correctly folded structure of proteins as chaperones. During viral infection, some HSPs can have an antiviral effect by inhibiting viral proliferation through interaction and activating immune pathways to protect the host cell. However, although the biological function of HSPs is to maintain the homeostasis of cells, some HSPs will also be hijacked by viruses to help their invasion, replication, and maturation, thereby increasing the chances of viral survival in unfavorable conditions inside the host cell. In this review, we summarize the roles of the heat shock protein family in various stages of viral infection and the potential uses of these proteins in antiviral therapy.

Net Optical Gain Coefficients of Cu+ and Tm3+ Single-Doped and Co-Doped Germanate Glasses.

Broadband tunable solid-state lasers continue to present challenges to scientists today. The gain medium is significant for realizing broadband tunable solid-state lasers. In this investigation, the optical gain performance for Tm3+ and Cu+ single-doped and co-doped germanate glasses with broadband emissions was studied via an amplified spontaneous emission (ASE) technique. It was found that the net optical gain coefficients (NOGCs) of Tm3+ single-doped glass were larger than those for Cu+ single-doped glass. When Tm3+ was introduced, the emission broadband width of Cu+-doped glass was effectively extended. Moreover, it was found that for the co-doped glass the NOGCs at the wavelengths for Tm3+ and Cu+ emissions were larger than those of Tm3+ and Cu+ single-doped glasses at the same wavelengths. In addition, the NOGC values of Tm3+ and Cu+ co-doped germanate glasses were of the same order of magnitude, and were maintained in a stable range at different wavelengths. These results indicate that the Tm3+ and Cu+ co-doped glasses studied may be a good candidate medium for broadband tunable solid-state lasers.

Turn-on fluorescence ferrous ions detection based on MnO2 nanosheets modified upconverion nanoparticles.

A turn on upconversion fluorescence probe based on the combination of ~32 nm NaYF4: Yb/Tm nanoparticles and MnO2 nanosheets has been established for rapid, sensitive detection of Fe2+ ions levels in aqueous solutions and serum. X-ray diffraction (XRD), transmission electron microscopy (TEM), absorption and emission spectra have been used to characterize the crystal structure, morphology and optical properties of the samples. MnO2 nanosheets on the surface of UCNPs act as a fluorescence quencher, resulting in the quenching of the blue fluorescence (with excitation/emission maximum of 980/476 nm) via fluorescence resonance energy transfer from upconversion nanoparticles to MnO2 nanosheets. With the adding of Fe2+, upconversion fluorescence of the nanocomposites recovers due to the reduction of MnO2 to Mn2+. Because of the low background of the probe offered by upconversion fluorescence, this probe can be used for detecting Fe2+ in aqueous solutions in the range of 0.1-22 µM with detection limit of 0.113 µM. The developed method has also been applied to detect 10 µM Fe2+ ions in serum with recoveries ranging from 97.6 to 105.3% for the five serum samples. Significantly, the probe shows fast response and stable signal, which is beneficial for long-time dynamic sensing. Thus, the proposed strategy holds great potential for disease diagnosis and treatment.

Fluorescence decay route of optical transition calculation for trivalent rare earth ions and its application for Er3+-doped NaYF4 phosphor.

Usually, the optical transition properties of trivalent rare earth (RE) ions in transparent hosts can be quantitatively investigated in the framework of Judd-Ofelt theory. A standard and commonly accepted calculation procedure based on the absorption spectrum has already been established. However, it is hard to assess the optical transition properties of trivalent RE ions doped in powdered and film materials owing to the difficulty in the absolute absorption spectrum measurements. In this work, we proposed a new route to calculate the Judd-Ofelt parameters of trivalent RE ion-doped materials in any morphological and shaped forms. In this method, the fluorescence decay values bridging the radiative transition rates and the Judd-Ofelt parameters were used. As an application example of the proposed Judd-Ofelt calculation method, the Judd-Ofelt parameters of Er3+ in NaYF4 were calculated via the proposed route, and it was found that the obtained results are in reasonable accordance with those derived from other routes. It was also proved that this proposed Judd-Ofelt calculation method is a practicable and effective route for evaluating optical transition properties of trivalent RE ions in non-transparent hosts as long as the fluorescence decay values can be measured.

Restoration and Calibration of Tilting Hyperspectral Super-Resolution Image.

Tilting sampling is a novel sampling mode for achieving a higher resolution of hyperspectral imagery. However, most studies on the tilting image have only focused on a single band, which loses the features of hyperspectral imagery. This study focuses on the restoration of tilting hyperspectral imagery and the practicality of its results. First, we reduced the huge data of tilting hyperspectral imagery by the p-value sparse matrix band selection method (pSMBS). Then, we restored the reduced imagery by optimal reciprocal cell combined modulation transfer function (MTF) method. Next, we built the relationship between the restored tilting image and the original normal image. We employed the least square method to solve the calibration equation for each band. Finally, the calibrated tilting image and original normal image were both classified by the unsupervised classification method (K-means) to confirm the practicality of calibrated tilting images in remote sensing applications. The results of classification demonstrate the optimal reciprocal cell combined MTF method can effectively restore the tilting image and the calibrated tiling image can be used in remote sensing applications. The restored and calibrated tilting image has a higher resolution and better spectral fidelity.

Influence of Er3+ concentration and Ln3+ on the Judd-Ofelt parameters in LnOCl (Ln = Y, La, Gd) phosphors.

The optical transition properties of trivalent rare earth (RE3+) doped luminescent materials have received extensive attention. The Judd-Ofelt theory is an effective tool for exploring the optical transition properties for the 4f-4f transitions of lanthanides. The aim of this work is to discover the effect of Er3+ concentration and different Ln3+ ions on the Judd-Ofelt parameters in LnOCl:Er3+ (Ln = Y, La, Gd) phosphors. Oxychloride LnOCl:Er3+ (Ln = Y, La, Gd) phosphors were produced via a single displacement reaction technique. The Judd-Ofelt calculation procedure for RE3+ doped powders was modified and then adopted to obtain the Judd-Ofelt parameters of Er3+ in the studied phosphors. Meanwhile, a new route for examining the Judd-Ofelt calculation quality was proposed and used. It was found that the Er3+ doping concentration slightly affects the optical transition properties of Er3+ in YOCl and LaOCl, but greatly affects the optical transition properties in GdOCl. Moreover, it was also found that the optical transition properties of Er3+ depend also on Ln3+ (Ln = Y, La, Gd) though the crystal structure of these compounds is similar. The Judd-Ofelt parameters of Er3+ are the smallest in LaOCl:Er3+, medium in YOCl:Er3+, but the biggest in GdOCl:Er3+ when the doping concentration is the same.

Cascade Synthesis from Cyclohexane to ϵ-Caprolactone by Visible-Light-Driven Photocatalysis Combined with Whole-Cell Biological Oxidation.

Cyclohexane was directly oxy-functionalised to ϵ-caprolactone through a cascade reaction sequence combining visible-light-driven photocatalysis with cyclohexanone monooxygenase (CHMO) whole-cell biocatalysis. Two available photocatalysts, Au-doped TiO2 (Au-TiO2 ) and graphitic carbonitride (g-C3 N4 ), were evaluated in the experiment and some optimisations to the cascade reaction were applied. In stepwise mode, the highest degree of conversion from cyclohexanol to ϵ-caprolactone can be up to 41 %, with use of g-C3 N4 . The cascade reaction from cyclohexane to ϵ-caprolactone is achievable under a light intensity of 149 µW cm-2 .

Enantioselective Sulfoxidation of Thioanisole by Cascading a Choline Oxidase and a Peroxygenase in the Presence of Natural Deep Eutectic Solvents.

A bienzymatic cascade for selective sulfoxidation is presented. The evolved recombinant peroxygenase from Agrocybe aegeritra catalyses the enantioselective sulfoxidation of thioanisole whereas the choline oxidase from Arthrobacter nicotianae provides the H2 O2 necessary via reductive activation of ambient oxygen. The reactions are performed in choline chloride-based deep eutectic solvents serving as co-solvent and stoichiometric reductant at the same time. Very promising product concentrations (up to 15 mM enantiopure sulfoxide) and catalyst performances (turnover numbers of 150,000 and 2100 for the peroxygenase and oxidase, respectively) have been achieved.

Multiple logic operations based on chemically triggered upconversion fluorescence switching.

The development of upconversion nanoparticles based logic systems, especially integrated logic systems is still a challenge until now. In this work, an upconversion nanocomposite system is developed and studied for the sensing abilities toward hydrion, hydroxyl ions, metal ions and anions (S2-, I-) by taking the advantages of turn-on and turn-off upconversion fluorescence switching response. Triggering by different kinds of ions, the upconversion system can act as a fluorescence switch due to the specific recognition abilities of Rhodamine 6G functionalized PEI for specific ions and the energy transfer process from upconversion nanoparticles to recognition molecules. Based on these results, multiple molecular logic gates, including single-input logic operation (YES, NOT), double-inputs logic operation (OR, AND, NOR, INHIBIT) and multiple-input integrative logic operation (INHIBIT+OR) are developed by employing hydrion, hydroxyl ions, metal ions and anions as inputs and the changes in the upconversion fluorescence intensity as output. The multiple logic operations are of great significance for the applications in biomedicine and molecular calculation.

Anomalous Temperature-Dependent Upconversion Luminescence of α-NaYF4:Yb³âº/Er³âº Nanocrystals Synthesized by a Microwave-Assisted Hydrothermal Method.

Yb³âº/Er³âºco-doped cubic-(α-) phase NaYF4 nanocrystals were prepared through a microwave- assisted hydrothermal method. Temperature-dependent upconversion luminescence (UCL) and sensing properties were systematically studied. It is interesting that anomalous temperature- dependent UCL behavior is observed. With increasing temperature (303-573 K), the UCL intensity of Er³âº does not quench monotonously but reaches a minimum around 483 K and then increases. However, it was found that the UCL spectra change in a different way with decreasing temperature (573-303 K) from the one measured with increasing temperature. The fluorescence intensity ratio of ²H11/2 --> 4I15/2 to 4S3/2 --> 4I15/2 at any measured temperature point remains almost constant in all measurement processes, indicating the consistency of temperature in each spectrum measurement at all temperature points regardless of the heating or the cooling process in our experiments. The results demonstrate that NaYF4:Yb³âº/Er³âº UC nanocrystal has good sensing stability and may have potential application in the nanoscale thermal sensor.

Development a scalable production process for truncated human papillomavirus type-6 L1 protein using WAVE Bioreactor and hollow fiber membrane.

Here, we describe a process for expression, purification, and characterization of truncated human papillomavirus type-6 (HPV-6) L1 virus-like particles (VLPs). The scalable cultivation process in a WAVE Bioreactor at the 10-L scale was optimized to express HPV-6 L1 VLPs using the baculovirus insect expression system. A hollow fiber membrane system was used for the integrated operation, including concentration, diafiltration, extraction, and clarification. The HPV-6 L1 protein was further purified by anion-exchange chromatography and hydrophobic chromatography. The HPV-6 L1 protein could self-assemble into VLPs with a diameter of approximately 50-60 nm after removal of the reductant dithiothreitol (DTT). The final purified HPV-6 L1 VLPs product was characterized to estimate yield and purity, and exceeds the requirements for pharmaceutical-grade VLP vaccine. Immunization of mice demonstrated that the vaccine could elicit high titer neutralizing antibodies in vivo. This study confirms the feasibility of producing pharmaceutical-grade HPV type-6 L1 VLPs on an industrial scale for clinical trials.

Permeability of exendin-4-loaded chitosan nanoparticles across MDCK cell monolayers and rat small intestine.

The purpose of this study was to investigate the permeability of exendin-4-loaded chitosan nanoparticles using the Madin Darby canine kidney (MDCK) cell monolayer as an in vitro model and the rat intestine as an ex vivo model of the human intestinal barrier. A series of formulations of sodium tripolyphosphate (TPP) and chitosan with different molecular weights and degrees of deacetylation was evaluated. The formulation consisting of 0.1% TPP and 0.2% chitosan (400 kDa, 95% degree of deacetylation), which gave optimized monodispersed particle size (303.1±10.36 nm), zeta potential (18.37±1.15 mV) and encapsulation efficiency (38.0±2.6%), was used for further analysis. After determining their biocompatibility, the transport potential of drug-loaded chitosan nanoparticles was evaluated and compared with free exendin-4 using both MDCK cell monolayers and different rat intestinal segments. Mechanisms underlying enhanced transport of exendin-4 in the cell model were also explored. Compared with free exendin-4, the absorption of optimized chitosan nanoparticles was enhanced by 4.7-fold in MDCK cell monolayers and by 2.0-2.78-fold in different rat intestinal segments, with no significant difference between the duodenum, jejunum and ileum. As supported by confocal laser scanning microscopic analysis, the lower enhancement of absorption in the intestine compared to the cell monolayer likely resulted from the chitosan nanoparticle-mediated opening of cellular tight junctions and not through intracellular transport. These findings suggest that the potential application of chitosan nanoparticles as delivery carriers of exendin-4 is limited and may need further modifications.

Negative effects of a disulfide bond mismatch in anti-rabies G protein single-chain antibody variable fragment FV57.

Rabies virus (RV) causes a fatal infectious disease requiring efficient post-exposure prophylaxis (PEP), which includes a rabies vaccine and rabies immunoglobulin (RIG). The single-chain antibody variable fragment (scFv), a small engineered antibody fragment derived from an antibody variable heavy chain and light chain, has the potential to replace the current application of RIG. In previous studies, we constructed and evaluated an anti-rabies virus G protein scFv (FV57) based on the monoclonal antibody CR57. Of the five cysteines in FV57, four are linked in intra-chain disulfide bonds (Cys-VH28/Cys-VH98 and Cys-VL16/Cys-VL84), and one is free (Cys-VL85). However, the thiol in Cys-VL85 neighboring Cys-VL84 in the CDR3 of the light chain is likely to mismatch with the thiol in Cys-VL16 during the renaturing process. In order to study effects of the mismatched disulfide bond, Cys-VL85 and Cys-VL84 of FV57 were mutated to serine to construct mutants FV57(VL85S) and FV57(VL84S). Furthermore, the disulfide bonds in the light chain of FV57, FV57(VL85S) and FV57(VL84S) were deleted by mutating Cys-VL16 to serine. All mutants were prepared and evaluated along with the original FV57. The results indicated that the mismatched disulfide bond of FV57 linking the light chain FR1 and CDR3 would confer deleterious negative effects on its activity against RV, likely due to spatial hindrance in the light chain CDR3. Moreover, avoidance of the disulfide bond mismatch provided an additional 30% protective efficacy against RV infection in the mouse RV challenge model. Thus, modifications of FV57 to eliminate the disulfide bond mismatch may provide a candidate therapeutic agent for effective PEP against rabies.

Sphere formation assay is not an effective method for cancer stem cell derivation and characterization from the Caco-2 colorectal cell line.

Although the existence of cancer stem cells (CSCs) has been demonstrated in colorectal cancer, further investigation is hindered by controversies over their surface markers. The sphere formation assay is widely used as in vitro method for derivation and characterization of CSCs based on the intrinsic self-renewal property of these cells. Isolated cancer cells that form tumorspheres are generally recognized as CSCs with self-renewal and tumorigenic capacities. In this study, colon spheres grown from Caco-2 cells in the sphere formation assay were separated from other differentiated cells and characterized. Compared with Caco-2 cells, the derived colon spheres lost several CSC properties. The colon spheres contained decreased levels of specific colorectal CSC surface markers as well as low levels of ATP-binding cassette (ABC) transporters typically overexpressed in CSCs, resulting in the near loss of their chemoresistance ability. Furthermore, cells that developed as colon spheres with strong self-renewal ability in vitro lost their tumorigenic capacity in vivo compared with Caco-2 cells, which could establish tumors in non-obese diabetic/severe-combined immunodeficient (NOD/SCID) mice. The results indicated that the Caco-2 cell derived colon spheres did not consist of colorectal CSCs. Thus, the well-accepted sphere formation assay may not be an effective method for CSC isolation and characterization from the Caco-2 colorectal cancer cell line.

Large-scale purification of pharmaceutical-grade plasmid DNA using tangential flow filtration and multi-step chromatography.

The demand for pharmaceutical-grade plasmid DNA in vaccine applications and gene therapy has been increasing in recent years. In the present study, a process consisting of alkaline lysis, tangential flow filtration, purification by anion exchange chromatography, hydrophobic interaction chromatography and size exclusion chromatography was developed. The final product met the requirements for pharmaceutical-grade plasmid DNA. The chromosomal DNA content was <1 µg/mg plasmid DNA, and RNA was not detectable by agarose gel electrophoresis. Moreover, the protein content was <2 µg/mg plasmid DNA, and the endotoxin content was <10 EU/mg plasmid DNA. The process was scaled up to yield 800 mg of pharmaceutical-grade plasmid DNA from approximately 2 kg of bacterial cell paste. The overall yield of the final plasmid DNA reached 48%. Therefore, we have established a rapid and efficient production process for pharmaceutical-grade plasmid DNA.