• Title/Summary/Keyword: cell yield

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Increased Refolding Yield of Disulfide Bond Bridged Fab-Toxin Homodimers by the Insertion of CH3 Domains

  • Song Jeong-Wha;Won Jae-Seon;Lee Yong-Chan;Choe Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1104-1110
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    • 2006
  • Recombinant antibody-toxin is a bifunctional protein that binds and kills a target cell expressing a specific antigen on the surface of the cell, and its structure is chimeric, in which a toxin is fused to an antigen-binding domain such as scFv or Fab. Divalent antibody-toxin molecules showed higher cytotoxicities against cancer cell lines than monovalent molecules. However, the yields of the divalent molecules were very low. In this study, we introduced the CH2, CH3, or CH2-CH3 (=Fc) domain of antibody in the middle of the Fab-toxin between the hinge region of human IgG1 and the toxin domain to increase the yield. The covalently bonded dimer could be formed by three disulfide bridges from cysteine residues in the hinge region. The molecule with the CH3 domain showed about 3-fold higher dimerization yield than previously constructed Fab-toxin molecules, while maintaining the cytotoxic activity comparable to that of scFv-toxin. However, the introduction of CH2 or Fc domain to the same position showed little effect on the dimerization yield. We also observed that the introduction of the CH3 region made it possible to form noncovalently associated dimer molecules.

Optimization of Host Animal Cell Culture Conditions to Produce Protein Using Recombinant Vaccinia Virus (재조합 백시니아 바이러스를 이용한 단백질 생산을 위한 숙주 동물세포의 배양 조건 최적화)

  • 이두훈;박정극
    • KSBB Journal
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    • v.11 no.4
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    • pp.438-444
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    • 1996
  • Using recombinant Vaccinia virus(vSC8) that express ${\beta}$-galactosidase, a model heterologous protein, conditions for virus and protein production were investigated in tissue culture flask. As host animal cells HeLa and HeLa S3 were used. It was demonstrated that cells infected during the exponential growth phase gave higher protein yield than those infected during the stationary growth phase and calf serum concentration after virus infection did not significantly alter protein yield. Pretreatment of cell layer with hypotonic solution enhanced the virus infectivity. Optimum cell growth and recombinant protein production was achieved at $37^{\circ}C$. But, during 2 hours of virus infection period incubation temperature must be lowered to 20∼$30^{\circ}C$ for maximum recombinant protein yield. To enhance virus replication, the effects of adrenal glucocorticoid hormone (Dexamethasone) and silkworm hemolymph were evaluated. Only dexamethasone increased about 20% of ${\beta}$-galactosidase yield in HeLa S3 cells when added with 10-7∼10-5M concentration 24 hours before infection.

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Studies on the Polysaccharide of Ganoderma lucidium Extract by Microorganism Fermentation (미생물 발효 영지버섯 추출액의 다당체에 관한 연구)

  • Hwang, Yu-Yeon;Chong, Myong-Soo;Kim, Hae-Ja;Lee, Ki-Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1506-1512
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    • 2007
  • In order to investigate the totale yield, the content of soluble polysaccharide and the others of FEW extract from the yeast fermentated Ganoderma lucidium by supersonic method, the yeast strain was inoculated after pretreatment and subsequently followed fermentation and supersonic extract. The main construction of the extract method is composed of the main glucose and together with the xylose, fucose, galactose and mannose. Results show that because of the generated lactic acid and ethanol, pH value of extract decreases and the safety as well as the preservation is improved. The extract yield, the total soluble polysaccharide, SOD-like activity, cytotoxic effect and growth inhibitory effect against cancer cell line are much higher in FE method than RE method, especially FEW3 extracts fermented during 24hrs. It is concluded that yeast fermentation makes the extract yield increase because of the cell disintegration, the useful ingredients of the germ body, the metabolic products, the insoluble ingredients due to the generation of ethanol, and the cell fragmentation caused by the supersonic waves vibration. Content of generated ethanol, total soluble polysaccharide and extract yield all increase during the fermentation time from 24 to 72 hours and the optimum fermentation condition is at $27^{\circ}C$ for 72 hours. The bitter taste and smell of the Ganoderma lucidium extract is diminished, fragrant-bitter taste and smell is generated so that the whole functional quality is improved.

Characteristics of Barkhausen Noise Properties and Hysteresis Loop on Tensile Stressed Rolled Steels

  • Kikuchi, Hiroaki;Ara, Katsuyuki;Kamada, Yasuhiro;Kobayashi, Satoru
    • Journal of Magnetics
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    • v.16 no.4
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    • pp.427-430
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    • 2011
  • The rolled steels for welded structure applied tensile stress have been examined by means of magnetic Barkhausen noise (MBN) method and of a physical parameter obtained from a hysteresis loop. The behaviors of MBN parameters and coercive force with tensile stress were discussed in relation to microstructure changes. There is no change in MBN parameters and coercive force below yield strength. The coercive force rises rapidly with tensile stress above yield strength. On the other hand, the rms voltage and the peak in averaged rms voltage take a maximum around yield strength and then decreases. The magnetomotive force at peak in the averaged rms voltage shows a minimum around yield strength. These phenomena are attributed to the combined effects of cell texture and dislocation density. In addition, the behaviors of MBN parameters around yield strength may be reflected by the localized changes in strain field due to the formation of dislocation tangles.

Genetic Evaluation of Somatic Cell Counts of Holstein Cattle in Zimbabwe

  • Mangwiro, F.K.;Mhlanga, F.N.;Dzama, K.;Makuza, S.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1347-1352
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    • 2000
  • The objectives of the study were to examine non-genetic factors that influence somatic cell counts in dairy cattle and to estimate the genetic parameters of somatic cell counts. A total of 34, 097-test day somatic cell count records were obtained from the Zimbabwe Dairy Services Association (ZDSA). The data were from 5, 615 Holstein daughters of 390 sires and 2, 541 dams tested between May 1994 and December 1998. First lactation cows contributed 22, 147 records to the data set, while 11, 950 records were from second and later parity cows. The model for analysis included fixed effects of month of calving, year of calving, stage of lactation, calving interval and test date. Milk yield and age on test day were fitted in the model as covariates. The additive genetic effects pertaining to cows, sires and dams and the residual error were the random effects. The Average Information Restricted Maximum Likelihood algorithm was used for analysis. The heritability of somatic cell scores was low at $0.027{\pm}0.013$ for parity one cows and $0.087{\pm}0.031$ for parity two and above. Repeatability estimates were $0.22{\pm}0.01$ and $0.30{\pm}0.01$ for the two lactation groups, respectively. Genetic and phenotypic correlations between the somatic cell scores and test day milk production were small and negative. It seems that there is no genetic link between somatic cell counts and milk yield in Holstein cattle in Zimbabwe. The results also seem to indicate that somatic cell count is a trait that is mainly governed by environmental factors.

Prediction of Watershed Erosion and Deposition Potentials (유역침식 및 퇴적 잠재능 예측모델 개발)

  • Son, Kwang-Ik
    • Journal of the Korean Society of Hazard Mitigation
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    • v.7 no.1 s.24
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    • pp.67-72
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    • 2007
  • A model for predicting potentials of land erosion and deposition over a natural basin was developed based on the mass balance principle. The program was developed based on sediment mass balance principle for each cell in a GIS. Sediment yield from a cell was estimated with RUSLE. The outflow sediment from a cell was calculated by multiplying the sediment yield of the cell by the sediment delivery ratio (SDR) of the cell. The outflow sediment from the upstream cell becomes the incoming sediment of the downstream cell. Therefore the erosion and deposition potential of each cell could be determined from the sediment mass balance i.e., the difference between the incoming and outflow of sediments of each cell. The developed model was validated by comparing the predicted sediment yields for three basins with measured data.

Protoplast Production from Sphacelaria fusca (Sphacelariales, Phaeophyceae) Using Commercial Enzymes

  • Avila-Peltroche, Jose;Won, Boo Yeon
    • Journal of Marine Bioscience and Biotechnology
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    • v.12 no.1
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    • pp.50-58
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    • 2020
  • Sphacelaria is a filamentous brown algal genus that can be epibiotic on macroalgae, marine plants, and sea turtles. Its important role in benthic ecosystems, exposure to different stressors (e.g., grazing), and use as a model organism make Sphacelaria ideal for assessing physiological responses of organisms to environmental inputs. Single-cell RNA sequencing is a powerful new probe for understanding environmental responses of organisms at the molecular (transcriptome) level, capable of delineating gene regulation in different cell types. In the case of plants, this technique requires protoplasts ("naked" plant cells). The existing protoplast isolation protocols for Sphacelaria use non-commercial enzymes and are low-yielding. This study is the first to report the production of protoplasts from Sphacelaria fusca (Hudson) S.F. Gray, using a combination of commercial enzymes, chelation, and osmolarity treatment. A simple combination of commercial enzymes (cellulase Onozuka RS, alginate lyase, and driselase) with chelation pretreatment and an increased osmolarity (2512 mOsm/L H2O) gave a protoplast yield of 15.08 ± 5.31 × 104 protoplasts/g fresh weight, with all the Sphacelaria cell types represented. Driselase had no crucial effect on the protoplast isolation. However, the increased osmolarity had a highly significant and positive effect on the protoplast isolation, and chelation pretreatment was essential for optimal protoplast yield. The protocol represents a significant step forward for studies on Sphacelaria by efficiently generating protoplasts suitable for cellular studies, including single-cell RNA sequencing and expression profiling.

Development of Useful Products Through Plant Cell Fusion and Culture of Populus spp.(II) (식물세포 배양 및 융합을 통한 유용물질 개발(II))

  • Kim, K.U.;Park, Y.G.;Choi, M.S.
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.160-165
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    • 1995
  • Anthocyanin formation in callus cultures using Populus alba ${\times}$ Populus glandulosa was evaluated on basal MS medium supplemented with various levels of growth regulators, sucrose and nitrate concentrations. The highest yield of anthocyanin from cultured cells was produced under 5% sucrose, 1/8 strength of nitrate(12.5% of basic concentration) and combination of 1.0 mg/l IAA with 2 mg/l BAP, respectively. The high anthocyanin producing cell line no. 11 was selected among 15 cell lines, showing over 80% cells contained anthocyanin producing cells. From these cells, the highly productive red protoplast was isolated and the highest protoplast yield, $6.7{\times}10^6$ was obtained in enzyme combination IV which is composed of 2.0% cellulase, 0.5% macerozyme and 0.1% pectolyase.

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Growth Prpmotion of Taxus brevifolia Cell Suspension Culture Using Conditioned Medium

  • Kim, Myung-Hwan;Chun, Su-Mwan;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.350-354
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    • 2000
  • The growth promotion of a Taxus brevifolia cell suspension culture was investigated using conditioning factors. The conditioning factors produced and secreted from cultured cells usually stimulate cell division and the production of secondary metabolites. Therefore, the effective incubation time for the optimal secretion of conditioning factors was firstly determined for the promotion of cell growth. Conditioned media obtained by cultivating for 2 and 5 days showed the promotion of initial cell growth during the early cell growth period. However, the positive effect of the conditioning factors on the initial cell growth did not continue because of the depletion of the medium nutrients. Accordingly, the addition of a carbon source to the conditioned medium prolonged the positive effect on the cell growth. The addition of sucrose to the conditioned medium resulted in the maximum cell density being reached 4 days earlier compared to the control group and an increased substrate yield.

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Improvement of Anthocyanin Encapsulation Efficiency into Yeast Cell by Plasmolysis, Ethanol, and Anthocyanin Concentration Using Response Surface Methodology

  • Dong, Lieu My;Hang, Hoang Thi Thuy;Tran, Nguyen Huyen Nguyet;Thuy, Dang Thi Kim
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.267-275
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    • 2020
  • Anthocyanins are antioxidant compounds susceptible to environmental factors. Anthocyanin encapsulation into yeast cells is a viable solution to overcome this problem. In this study, the optimal factors for anthocyanin encapsulation were investigated, including anthocyanin concentration, plasmolysis contraction agent, and ethanol concentration, and response surface methodology was evaluated, for the first time. Anthocyanin from Hibiscus sabdariffa L. flowers was encapsulated into Saccharomyces cerevisiae using plasmolysis contraction agent (B: 3%-20% w/v), ethanol concentration (C: 3%-20% v/v), and anthocyanin concentration (A: 0.15-0.45 g/ml). The encapsulation yield and anthocyanin loss rate were determined using a spectrometer (520 nm), and color stability evaluation of the capsules was performed at 80℃ for 30 min. The results of the study showed that these factors have a significant impact on the encapsulation of anthocyanin, in which ethanol agents have the highest encapsulation yield compared to other factors in the study. Statistical analysis shows that the independent variables (A, B, C), their squares (A2, B2, C2), and the interaction between B and C have a significant effect on the encapsulation yield. The optimized factors were anthocyanin, 0.25 g/ml; NaCl, 9.5% (w/v); and ethanol, 11% (v/v) with an encapsulation yield of 36.56% ± 0.55% and anthocyanin loss rate of 15.15% ± 0.98%; This is consistent with the expected encapsulation yield of 35.46% and loss rate of 13.2%.