| 1. |
- Nilsang, Suthasinee, et al.
(författare)
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Three-dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles
- 2008
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Ingår i: Biotechnology progress (Print). - Malden, MA : Wiley-Blackwell Publishing Inc.. - 8756-7938 .- 1520-6033. ; 24:5, s. 1122-1131
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Tidskriftsartikel (refereegranskat)abstract
- Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAXtrade mark) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on GlutaMAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mug mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production. © 2008 American Institute of Chemical Engineers.
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| 2. |
- Basheer, Shabana, et al.
(författare)
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A membrane protein based biosensor: Use of a phosphate - H+ symporter membrane protein (Pho84) in the sensing of phosphate ions.
- 2011
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Ingår i: Biosensors & bioelectronics. - : Elsevier. - 0956-5663 .- 1873-4235. ; 27:1, s. 58-63
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Tidskriftsartikel (refereegranskat)abstract
- A label free biosensor for direct detection of inorganic phosphate based on potential-step capacitance measurements has been developed. The high-affinity Pho84 plasma membrane phosphate/proton symporter of Saccharomyces cerevisiae was used as a sensing element. Heterologously expressed and purified Pho84 protein was immobilized on a self-assembled monolayer (SAM) on a capacitance electrode. Changes in capacitance were recorded upon exposure to phosphate compared to the control substance, phosphate analogue methylphosphonate. Hence, even without the explicit use of lipid membranes, the Pho84 membrane protein could retain its capacity of selective substrate binding, with a phosphate detection limit in the range of the apparent in vivo K(m). A linear increase in capacitance was monitored in the phosphate concentration range of 5-25 mu M. The analytical response of the capacitive biosensor is in agreement with that the transporter undergoes significant conformational changes upon exposure to inorganic phosphate, while exposure to the analogue only causes minor responses.
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| 3. |
- Håkansson, Torbjörn, et al.
(författare)
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Sulphate reducing bacteria to precipitate mercury after electrokinetic soil remediation
- 2008
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Ingår i: International journal of environmental science and technology. - 1735-1472 .- 1735-2630. ; 5:2, s. 267-274
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Tidskriftsartikel (refereegranskat)abstract
- Combined treatment with electroremediation and sulphate reducing bacteria (SRB) was tested in laboratory and pilot scale. The contaminated soil came from a chlor-alkali factory and contained about 100 mg/kg Hg. Iodide/iodine complexing agent was used to mobilize mercury. Mercury iodide complexes were moved to the anode solution using an electric field. The anode solution was then mixed with hydrogen sulphide (H2S) containing water, causing precipitation of mercury sulphide. The H2S was produced at site by a SRB reactor. Precipitation problems arising from the nature of the anode solution were expected, since this solution is highly acidic, very oxidised and may contain iodide/iodine that strongly complexes mercury and can hinder mercury sulphide precipitation. Mercury concentrations in the anode solution were up to 65.7 mg/L (field) and 15.4 mg/L (lab. scale). Reduction of mercury in the water was >93% at all times. Iodide did not hinder the process: Nonetheless, in the lab system, iodide concentration was high in the anode solution but mercury reduction was > 99.9%. The redox potential was sufficiently low for HgS precipitation during the experiments, except for a short period, when the mercury removal decreased to 94%. Sulphate reducing bacteria are shown as a viable tool for the treatment of mercury contaminated, acidic, oxidative, iodide containing water, such as that produced by electrokinetic remediation. A second SRB step or other water treatment is required to reduce the mercury concentration to environmentally acceptable levels. Redox. potential is the most sensitive factor in the system.
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| 4. |
- Punzi, Marisa, et al.
(författare)
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Combined anaerobic-ozonation process for treatment of textile wastewater : Removal of acute toxicity and mutagenicity
- 2015
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Ingår i: Journal of Hazardous Materials. - : Elsevier BV. - 0304-3894 .- 1873-3336. ; 292, s. 52-60
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Tidskriftsartikel (refereegranskat)abstract
- A novel set up composed of an anaerobic biofilm reactor followed by ozonation was used for treatment of artificial and real textile effluents containing azo dyes. The biological treatment efficiently removed chemical oxygen demand and color. Ozonation further reduced the organic content of the effluents and was very important for the degradation of aromatic compounds, as shown by the reduction of UV absorbance. The acute toxicity toward Vibrio fischeri and the shrimp Artemia salina increased after the biological treatment. No toxicity was detected after ozonation with the exception of the synthetic effluent containing the highest concentration, 1. g/l, of the azo dye Remazol Red. Both untreated and biologically treated textile effluents were found to have mutagenic effects. The mutagenicity increased even further after 1. min of ozonation. No mutagenicity was however detected in the effluents subjected to longer exposure to ozone. The results of this study suggest that the use of ozonation as short post-treatment after a biological process can be beneficial for the degradation of recalcitrant compounds and the removal of toxicity of textile wastewater. However, monitoring of toxicity and especially mutagenicity is crucial and should always be used to assess the success of a treatment strategy. © 2015 Elsevier B.V.
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| 5. |
- Punzi, Marisa, et al.
(författare)
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Degradation of a textile azo dye using biological treatment followed by photo-Fenton oxidation : Evaluation of toxicity and microbial community structure
- 2015
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Ingår i: Chemical Engineering Journal. - : Elsevier BV. - 1385-8947 .- 1873-3212. ; 270, s. 290-299
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Tidskriftsartikel (refereegranskat)abstract
- Many commercial dye preparations are cocktails of active dyes and various by-products that are recalcitrant to biological degradation and end up in significant amounts in the effluent after the dyeing process. Conventional wastewater treatment processes are not able to degrade such compounds and detoxify the effluent, thus alternative treatments should be developed.In our work we suggest to use photo-Fenton oxidation as post-treatment after an anaerobic biofilm process, in a way to minimize the reagents needed. This process was used for treatment of synthetic textile wastewater containing the commercial azo dyestuff Remazol Red, starch and sodium chloride. The treated textile effluent had COD lower than 18. mg/l even when using initial Fenton reagents concentration as low as 1. mM ferrous ions and 10. mM hydrogen peroxide. The acute toxicity was higher in the biologically treated than in the untreated effluent. Photo-Fenton oxidation successfully reduced the toxicity and the final effluent was non-toxic to Artemia salina and Microtox, with the exception of the effluent containing high concentration of sodium chloride, which was moderately toxic to Microtox. For the first time the presence of algae was detected in a reactor treating textile wastewater using denaturing gradient gel electrophoresis (DGGE); bacteria and fungi were also abundant.The results of this study suggest that using advanced oxidation after biological treatment is an effective way to degrade the organic compounds and remove toxicity from textile effluents.
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| 6. |
- Ertürk, Gizem, et al.
(författare)
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Highly sensitive detection and quantification of the secreted bacterial benevolence factor RoxP using a capacitive biosensor : A possible early detection system for oxidative skin diseases
- 2018
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Ingår i: PLOS ONE. - : Public Library of Science. - 1932-6203. ; 13:3
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Tidskriftsartikel (refereegranskat)abstract
- The impact of the microbiota on our health is rapidly gaining interest. While several bacteria have been associated with disease, and others being indicated as having a probiotic effect, the individual biomolecules behind these alterations are often not known. A major problem in the study of these factors in vivo is their low abundance in complex environments. We recently identified the first secreted bacterial antioxidant protein, RoxP, from the skin commensal Propionibacterium acnes, suggesting its relevance for maintaining the redox homeostasis on the skin. In order to study the effect, and prevalence, of RoxP in vivo, a capacitive biosensor with a recognition surface based on molecular imprinting was used to detect RoxP on skin in vivo. In vitro analyses demonstrated the ability to detect and quantify RoxP in a concentration range of 1 x 10(-13) M to 1 x 10(-8) M from human skin swabs; with a limit of detection of 2.5 x 10(-19) M in buffer systems. Further, the biosensor was highly selective, not responding to any other secreted protein from P. acnes. Thus, it was possible to demonstrate the presence, and quantity, of RoxP on human skin. Therefore, the developed biosensor is a very promising tool for the detection of RoxP from clinical samples, offering a rapid, cost-effective and sensitive means of detecting low-abundant bacterial proteins in vivo in complex milieus.
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| 7. |
- Kumar, Ashok, et al.
(författare)
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Affinity binding of cells to cryogel adsorbents with immobilized specific ligands : effect of ligand coupling and matrix architecture
- 2005
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Ingår i: Journal of Molecular Recognition. - : Wiley. - 0952-3499 .- 1099-1352. ; 18:1, s. 84-93
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Tidskriftsartikel (refereegranskat)abstract
- The capture of human acute myeloid leukemia KG-1 cells expressing the CD34 surface antigen and the fractionation of human blood lymphocytes were evaluated on polyvinyl alcohol (PVA)-cryogel beads and dimethyl acrylamide (DMAAm) monolithic cryogel with immobilized protein A. The affinity ligand (protein A) was chemically coupled to the reactive PVA-cryogel beads and epoxy-derivatized monolithic cryogels through different immobilization techniques and the binding efficiency of the cell surface receptors specific antibody-labeled cells to the gels/beads was determined. The binding of cells to monolithic cryogel was higher (90-95%) compared with cryogel beads (76%). B-lymphocytes, which bound to the protein A-cryogel beads, were separated from T-lymphocytes with yields for the two cell types 74 and 85%, respectively. About 91% of the bound B-cells could be recovered without significantly impairing their viability. Our results show differences in the percentage of cell-binding to the immunosorbents caused by ligand density, flow shear forces and bond strength between the cells and the affinity surface once distinct chemical coupling of protein A, size of beads, sequence of antibody binding to protein A adsorbents, morphology and geometry of surface matrices were compared.
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| 8. |
- Lacayo, Martha, et al.
(författare)
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A toxaphene-degrading bacterium related to Enterobacter cloacae, strain D1 isolated from aged contaminated soil in Nicaragua
- 2005
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Ingår i: Systematic and Applied Microbiology. - : Elsevier BV. - 0723-2020 .- 1618-0984. ; 28:7, s. 632-639
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Tidskriftsartikel (refereegranskat)abstract
- Enterobacter sp. strain D1 is a facultative anaerobic, Gram-negative heterotrophic bacterium isolated from toxaphene-contaminated soil. This organism was identified and characterized through phylogenetic and taxonomic studies. Based on 16S rDNA analysis, the strain D1 was clustered closely with the species Enterobacter cloacae subsp. dissolvens (LMG 2683) and E. cloacae (ATCC 13047T). Strain D1 resembled these E. cloacae strains with respect to various biochemical and nutritional characteristics, but also exhibited differences. Moreover, strain D1 is able to grow and survive with toxaphene supplied in the medium in the range 3-96 mg/L. Amongst the chemical components of toxaphene, octachlorocamphenes, nonachlorobornanes and decachlorobornanes were seen to be rapidly metabolized, although levels of hexachlorocamphenes and heptachlorobornanes were found to be slowly degraded, and subsequently accumulated during the last stage of the cultivation.
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| 9. |
- Lacayo, Martha, et al.
(författare)
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Degradation of toxaphene by Bjerkandera sp. strain BOL13 using waste biomass as a cosubstrate
- 2006
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Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 0175-7598 .- 1432-0614. ; 71:4, s. 549-554
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Tidskriftsartikel (refereegranskat)abstract
- The white-rot fungus Bjerkandera sp. strain BOL13 was capable of degrading toxaphene when supplied with wood chips, wheat husk or cane molasses as cosubstrates in batch culture experiments. Approximately 85% of toxaphene was removed when wheat husk was the main substrate. The production of lignin peroxidase was only stimulated when wheat husk was present in the liquid medium. Although xylanase was always detected, wheat husk supported the highest xylanase production. A negligible amount of beta-glucosidase and cellulase were found in the batch culture medium. To the best of our knowledge, this is the first reported case of toxaphene degradation by white-rot fungi.
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| 10. |
- Lacayo-Romero, Martha, et al.
(författare)
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Degradation of toxaphene in aged and freshly contaminated soil
- 2006
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Ingår i: Chemosphere. - : Elsevier BV. - 0045-6535 .- 1879-1298. ; 63:4, s. 609-615
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Tidskriftsartikel (refereegranskat)abstract
- Degradation of toxaphene in soil from both newly contaminated (from Sweden) and aged spills (from Nicaragua) were studied. The newly contaminated soil contained approximately 11 mg kg(-1) toxaphene while the aged Nicaraguan soil contained approximately 100 mg kg(-1). Degradation was studied in anaerobic bioreactors, some of which were supplied with lactic acid and others with Triton X-114. In this study we found that the lower isomers Parlar 11, 12 were degraded while the concentration of isomer Parlar 15 increased. This supported an earlier evaluation which indicated that less chlorinated isomers are formed from more heavily isomers. Lactic acid when added to the soil, interfere with the degradation of toxaphene. Lactic acid was added; several isomers appeared to degrade rather slowly in newly contaminated Swedish soil. The Swedish soil, without any external carbon source, showed the slowest degradation rate of all the compounds studied. When Triton X-114 at 0.4 mM was added, the degradation rate of the compounds increased. This study illustrates that biodegradation of toxaphene is a complex process and several parameters have to be taken into consideration. Degradation of persistent pollutants in the environment using biotechnology is dependent on bioavailability, carbon sources and formation of metabolites.
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