Very Strong But Reversible Immobilization of Enzymes on Supports Coated With
Cesar Mateo, Benevides C. C. Pessela, Manuel Fuentes, Rodrigo Torres, Claudia Ortiz, Fernando L pez-Gallego, Lorena Betancor, Noelia Alonso-Morales, Jose M. Guisan, and Roberto Fernandez-Lafuente In this chapter, the properties of tailor-made anionic exchanger resins based on films of large polyethylenimine polymers e.g., molecular weight 25,000 as supports for strong but reversible immobilization of proteins is shown. The polymer is completely coated, via covalent immobilization, the surface...
B
The matrix is activated and precoupled with an affinity ligand. The target enzyme is added. The enzyme is conjugated to another molecule, which in turn has affinity toward a matrix. The bioconjugate can be prepared by chemical cross-linking or obtained as a fusion protein. The enzyme is not subjected to harsh conditions. If the affinity ligand is a substrate product ana logue, the chances of enzyme dissociating from the affinity ligand are high. A more generic approach is possible for preparing...
Info Ptf
Fig. 7. Effect of the presence of lactose on the inactivation of derivative during the incubation with aldehyde-dextran. The assay was carried out in Novo buffer, pH 6.5 at 25 C. Values were reproduced in two separate experiments. All other conditions are detailed in Subheadings 2. and 3. derivative cross-linked with aldehyde-dextran incubated in the presence of 1 M lactose. derivative cross-linked with aldehyde-dextran. Fig. 8. Functional inactivation of optimal cross-linked derivative....
Info Tjk
1. Suggested ratios are shown in Table 1. However, other ratios, as well as other precursors, can be used. It should be taken into account that the higher the TMOS proportion, the faster the subsequent polymerization process. 2. Hydrolysis time depends on each case and should be optimized. 3. At basic pH, the condensation is favored. The appropriate basic pH will depend on the enzyme performance not too high to inactivate it . The buffer composition is not particularly important. However, NH4...
Info Njp
Fig. 5. 12 SDS-PAGE analysis of the P-galactosidase immobilized on boronic-epoxy-Sepabeads. Lane 1 molecular weight marker lane 2, desorption of the protein subunits not covalently immobilized on Sepabeads-boronic-epoxy support lane 3, desorption of proteins from optimal derivative and cross-linked with polyaldehyde-dextran. Arrows indicate the position of the P-galactosidase. Experiments were carried out in triplicate. All other specifications were described in Subheadings 2. and 3. However,...
Stability of Immobilized Enzymes on Polymeric Supports
3.8.1. Stability of Immobilized Multimeric Enzymes Immobilized on Ionic Polymer Coated Supports Invertase from Saccharomyces cerevisiae was immobilized in 5 mM sodium phosphate, pH 7.0, on DEAE-agarose or PEI supports see Subheading 3.3. , with an immobilization yield higher than 90 in both cases, after only 5 min of enzyme-support contact. The activity was fully preserved during the immobilization Fig. 8. Desorption of invertase from S. cerevisae adsorbed on different anion exchanging supports...
Covalent Immobilization of Glycosylated Proteins Previously Adsorbed Through
Starting from electrodes as in Subheading 3.4., boronic acids are incorporated into SAMs to reversibly adsorb glycosylated proteins through their carbohydrate moiety. A mixed monolayer of boronates and epoxy groups makes the adsorption irreversible. Basically, the aminophenylboronic groups promote and accelerate the covalent attachment of glycoproteins through the epoxy groups. 3.7.1. Amidation of the Thioctic Acid With Aminophenyl Boronic Acid and Epoxy Groups 1. Incubate overnight the gold...
Info Ddg
binding domain CBD -fi-galactosidase fusion protein on cellulosic supports resulted in a fairly efficient reusable biocatalyst 28 . Recently, combinatorial peptide synthesis has made it possible to obtain peptides as affinity ligands with a predetermined activity 5 . Such peptide affinity ligands, in principle, may also be used for affinity immobilization of enzymes since it will be possible to screen such ligands for binding constants in the range desirable for affinity immobilization. In this...
Info Uxp
Note The numbers express the amount of detergent necessary to desorb 100 of lipase from the support. Note The numbers express the amount of detergent necessary to desorb 100 of lipase from the support. evant conditions, it is convenient that the enzyme can, after enzyme deactivation, be desorbed from the support in order to reload the support with fresh enzyme and take advantage of the reversibility of the technique. Table 1 shows that the adsorbed lipases on octadecyl-Sepabeads or...
Preparation of Optical Biosensor Using Immobilized Microalgae 1819
3.2.1. Preparation of Microalgal Membrane 1. Cultivate the microalgae for example Chlorella vulgaris strain CCAP 211 12 in sterile culture medium. 2. To induce high AP activity, centrifuge the algal cultures and resuspend the pellets in phosphate-free medium for 25 d, which corresponds to maximal enzyme activity. But for the biosensors based on chlorophyll fluorescence measurement cultivate the algae under continuous mode see Note 4 . 3. Filter 2 mL of the resulting aliquot on a glass...
Stabilization of New Imprint Property of Glucose Oxidase in Pure Aqueous Medium
Bioimprinting is a well-known strategy to manipulate the catalytic properties of the enzymes. However, the lack of expression of the newly acquired imprinted property by an enzyme in aqueous surrounding is the chief limitation that restricts the application of this technique. In this chapter we used a combinatorial approach which amalgamates the benefits offered by traditional cross-link-immobilization technique rigidity and stability and bioimprinting approach induced tailor-made catalytic...
A
Fig. 2. Mechanism of the proteins immobilization on chelate supports. The IDA-Me supports are used to immobilize the proteins after they are incubated in the presence of mercaptoethanol to block the remaining epoxy groups and to release the Me2 from the support. Fig. 3. Selective adsorption of poly-His-tagged proteins on lowly activated IDA-Co2 epoxy supports. Fig. 3. Selective adsorption of poly-His-tagged proteins on lowly activated IDA-Co2 epoxy supports. Here, we present the preparation and...
Immobilization of LAsparaginase Into PolylactideCoglycolide Nanospheres
2.1.1. Preparation of i-Asparaginase-ioaded Nanospheres 1. L-Asparaginase from E. coli, Sigma, St. Louis, MO . 2. Polylactic-coglycolic acid 50 50 PLGA Resomer RG 503H, MW 34000 Da from Boehringer Ingelheim, Ingelheim, Germany see Note 1 . 3. Ethyl acetate Panreac Qu mica, Barcelona, Spain 4. Polyvinyl alcohol PVA MW 30,000-70,000, Sigma . 2.1.2. Characterization of i-Asparaginase-ioaded Nanospheres 1. Sodium hydroxide NaOH Panreac Qu mica . 2. Sodium dodecyl sulphate SDS Sigma . 3. 66...
Design of Smart Biocatalysts
Immobilization of Enzymes on Smart Polymers Ipsita Roy and Munishwar N. Gupta Smart polymers are water-soluble polymers that can be precipitated by an appropriate stimulus such as change of pH, ionic strength, temperature, or addition of a chemical species. Such polymers occur naturally e.g., alginate, chitosan but can also be synthesized chemically e.g., methyl methacrylate polymers available commercially as Eudragit . Linking of an enzyme to these polymers by noncovalent or covalent methods...
Ap
AP activity can easily be measured by measuring the MUF production at fluorescence emission of 460 nm under excitation light of 350 nm. The second type biosensor however, is based on algal chlorophyll fluorescence measurement at excitation of 482 nm and emission of 683 nm. 5. The substrate MUP was placed in the Tris buffer solution 0.1 M, pH 8.5 containing 1 mM MgCl2 as enzyme activator, and AP activity was measured in terms of fluorescence intensity. 6. AP activity can also be measured with...
Info Vno
effective for use in bioreactors, they may not be desirable in open environmental applications. The pore size of the support applied to a bioreactor process should be much smaller than the encapsulated cell to avoid or minimize cell release and washout from the bioreactor. Thus, cells remain inside the support while substrates and nutrients can diffuse in and products can diffuse out. In field applications, however, supports with high porosity are desirable because they provide high cell mass...
Info Rxo
nescence and therefore any interference results in effects observable as a loss of luminescence. Recently published applications of the luminescence biosensor exploiting immobilized cells are summarized in Table 1. Because the entire metabolism involved in bioluminescence is intracellular, effects on bioluminescence are likely to occur as a response to toxic compounds available in a form, which can be transported into the cell. As a result of the transport processes, correlation between...
Info Yoo
Fig. 11. Desorption of noncovalently immobilized protein subunits from the support in the prepared derivatives. Desorption was carried out by boiling the derivatives in the presence of SDS. Lane 1, crude extract lane 2, fraction of purified enzyme lane 3, enzyme immobilized on agarose-glutaraldehyde poorly activated 1 h of immobilization lane 4, enzyme immobilized on agarose-glutaraldehyde highly activated 24 h of immobilization lane 5, previous derivative modified with polyaldehyde-dextrans....
Preparation of Activated Solid Phases 1
The preparation of mercaptohydroxypropyl ether agarose gel thiol-agarose is carried out essentially as described by Axen et al. 9 . In this method the agarose beads Sepharose 4B are first reacted with epichlorohydrin in an alkaline medium. The oxirane groups thus formed are then converted with sodium thiosulfate to gel-bound thiosulfate groups Bunte-salt , which are finally reduced with DTT to thiol groups see Fig. 4 . 1. Suspend 15 g suction-dried Sepharose 4B in 15 mL of 1 MNaOH. 2. Add...
Immobilization of Enzymes for Use in Organic Media
When enzymes are to be used in organic media, noncovalent immobilization methods are often used because enzymes in general are insoluble in those media. Usually, immobilization increases the observed catalytic activity drastically as compared with nonimmobilized enzyme powder. The main reason is that immobilization spreads the enzyme on a relatively large area, which facilitates the mass transfer of substrates and products. The morphology of the support material is thus of crucial importance....
SinglePoint Attachment Binding Studies
3.2.1. Calorimetric Titration Study see Notes 6-7 1. Load the immobilized protein 0.25 g into the calorimeter titration cell and add 1 mL of a suitable buffer for example acetate buffer 50 mM, pH 5.0 . 2. Prepare 10 mL of 3'-CMP inhibitor solution 1 mMin the same buffer 10 immoles . 3. Pump the inhibitor solution at a constant rate ranging from 0.07 to 0.3 mL min until the heat effect decreases to baseline values about 20 min . 4. Prepare a second sample with the same amount of supporting...
Minimal Chemical Modification Minimal Physical Interactions Absence Of Steric
Fig. 8. Immobilization of an enzyme without altering its properties. 8. Enzyme Immobilization Still a Fascinating Challenge Thousands of protocols for enzyme immobilization have been reported over the past 30 yr. On their surfaces, enzymes have very different structural moieties that are able to interact i.e., adsorption covalent attachment with such activated supports as nucleophilic residues, hydrophobic pockets, carboxylic groups, regions with net-positive charge, and regions with...
Immobilized Enzymes for Biomedical Applications
Amaia Esquisabel, Rosa Mar a Hern ndez, Alicia Rodr guez Gasc n, and Jos Luis Pedraz Immobilized enzymes were first applied in the biomedical field aiming to detect bioactive substances or to treat a disease condition. This chapter presents two approaches used for the immobilization of l-asparaginase intended for the treatment of leukemia, based on the entrapment of this enzyme in a matrix. The particulate drug carriers described in this chapter are liposomes and nanoparticles, even though the...
Immobilization of Enzymes for Use in Supercritical Fluids
Pedro Lozano, Teresa de Diego, and Jos L. Iborra Supercritical fluids SCFs are environmentally benign solvents that enable efficient ability to dissolve and or transport many hydrophobic compounds. Because the solvent properties of SCFs can be adjusted by changing either the pressure or the temperature, they are widely used in a industrial extractive clean processes. For enzymatic biotransformation, several criteria must be considered to select an SCF as the reaction medium, such as the...
OneStep Purification Immobilization and Stabilization of PolyHistidineTagged
Cesar Mateo, Benevides C. C. Pessela, Valeria Grazu, Rodrigo Torres, Fernando Lopez-Gallego, Jose M. Guisan, and Roberto Fernandez-Lafuente In this chapter, the combined use of the selectivity of metal chelate affinity chromatography with the capacity of epoxy supports to immobilize poly-His-tagged proteins via multipoint covalent attachment is shown. This has been achieved by designing tailor-made chelate-epoxy supports. In order to selectively adsorb the poly-His-tagged proteins, a very small...
TaylorCouette Vortex Flow in Enzymatic Reactors
Roberto Campos Giordano and Raquel de Lima Camargo Giordano Taylor vortex flow reactors VFRs are especially useful when dealing with fragile biocatalysts, in view of the low-shear stress that is a characteristic of this flow pattern. Hence, they may be an interesting solution for reactions with immobilized enzymes. This chapter presents some basic features of this reactor. It is not intended to provide a review of the extensive literature on Taylor vortex flow reactors rather it includes some...
Bioluminescence in Immobilized Cells for Biomass Detection and Biosensor
Marian Navratil, Juraj Svitel, and Peter Gemeiner Bioluminescence measurements have become extremely popular because of good sensitivity and the ability to quantitate a wide variety of analytes. Only recently have these measurements gained particular interest in relation to the study of gene expression and regulation. The firefly luciferase system is the most frequently used bioluminescent reaction, in which adenosine triphosphate ATP is used to generate light. ATP is often used as a measure of...
Chemical Methods
The most extensively studied of the insolubilization techniques is the formation of covalent bonds between the enzymes and the support matrix. This is the retention of the enzyme on support surfaces by covalent bonding between functional groups on the enzyme and those on the support surface. The active site of the Fig. 1. Schematic representation of two occlusion methods for enzyme immobilization polymerization in PVA with styrilpirydinium groups PVA-SbQ A and enzyme entrapment in a sol-gel...
Eupergit C For Pga Immobilized
Fig. 4. Blockage of remainder epoxy groups with different compounds reactive with 4. Assay the enzyme activity of the reference solution using the same time intervals as in step 3. The immobilization process is finished when the activity of the supernatant is zero. 3.3. Desorption of Proteins Noncovalently Immobilized on the Support 1. 2.5 mL Enzymatic suspension were taken and dried by filtration under vacuum filter. 2. The dried support was resuspended in 2.5 mL of distilled water. The...
Immobilization of LAsparaginase Into Liposomes
2.2.1. Preparation of i-Asparaginase-ioaded iiposomes 1. Egg phospatidylcholine, cholesterol, and stearylamine Sigma . 2.2.2. Characterization of L-Asparaginase-Loaded Liposomes 3. 66 mMPhosphate buffer, pH 7.4. 6. 50 mM Tris buffer, pH 8.45 with 0.1 MHCl. 7. a-Ketoglutarate-sodium, L-glutamic oxaloacetate transaminase, L-malic dehydrogenase, NADH, and L-asparagine Sigma .
M
Fig. 1. The basic bioimprinting strategy. modified biomolecule is placed in aqueous medium again then the imprinted active site will revert back to its native conformation and cease to demonstrate the acquired imprinted property. To restrict this flipping back of a modified active site to its native and thermodynamically favorable conformation we developed the combinatorial cross-linked imprinted protein approach we termed it CLIP . The methodology is depicted in Fig. 2. Here, glucose oxidase...
o
Fig. 1. Chemical modification for achieving a more stable immobilized preparation. 3. Glycidol 2,3-epoxy propanol was purchased from Sigma-Aldrich St. Louis, MO see Note 2 . 4. Support oxidation solution 0.1 Msodium periodate in water. 5. Protein immobilization buffer 0.1 Mbicarbonate buffer, pH 10.0 see Note 3 . 1. Add 5 mL of 11 mg mL soluble enzyme to 45 mL of protein amination solution see Note 4 . 2. Stir gently for 90 min at 25 C and then add 10 mL of a 0.5 M hydroxylamine solution, pH...
Info Eqn
Fig. 6. Kinetic resolution of rac-6 catalyzed by lipases. Fig. 6. Kinetic resolution of rac-6 catalyzed by lipases. Fig. 7. Kinetic resolution of rac-9 catalyzed by lipases. Fig. 7. Kinetic resolution of rac-9 catalyzed by lipases. 1. n-Butyltrimethoxysilane ABCR, Germany . 2. Isobutyltrimethoxysilane Lancaster, UK . 3. Tween-80 poly-oxyethylene 20 sorbitan monooleate Fluka . 6. Methyl-P-cyclodextrin Aldrich . 8. Polyvinyl alcohol PVA molecular weight mol wt 15000 Merck, Germany . The lipases...
Notes Moi
1. Sterile mineral medium composition KNO3 25g, MgSO4 7H2O 10 g, KH2PO4 4 g, K2HPO4 1 g, FeSO4 7H2O 1 g, H3BO3 2.86 g, MnCl2 4H2O 1.81 g, ZnSO4 7H2O 0.11 g, CuSO4 5H2O 0.09 g, Na2MoO4 0.021 g L. Axenic Chlorella cultures were incubated for 5-6 d under continuous agitation 150 rpm , light intensity of 31.8 W m2 , and at 27-30 C. 2. Azospirillum brasilense strain Cd DMS 1843 was grown in liquid nutrient broth Sigma or M9 nitrogen-free medium composition g L Na2HPO4 6 g, KH2PO4 3 g, NaCl 1.5 g,...
Info Jpj
oped a special cross-linker based on dextran a mixture of glucose oligomers . The modified dextran is too large to enter the active site hence, in those cases where glutaraldehyde caused loss of activity, active CLEAs were obtained using this large molecule as a cross-linking agent. Because the enzyme molecules are packed together in a small volume as compared with the free protein in solution, one might expect mass-transport limitations with fast reactions. However, on the time scale of the...
Immobilization of Enzymes on Magnetic Particles
Martina Koneracka, Peter Kopcansky, Milan Timko, Chenyl Nynitapal Ramchand, Zainul M. Saiyed, Michael Trevan, Magnetic particles have been increasingly used as carriers for binding proteins, enzymes, and drugs. Such immobilization procedures for proteins, enzymes, antibodies, and other biologically active compounds have a major impact in different areas of biomedicine and biotechnology. The immobilized biomolecules can be used directly for a bioassay or as affinity ligands to capture or modify...
I111111
Fig. 3. Effect of ionic strength in the adsorption of lipase from R. niveus to octyl-agar-ose. 1 mL Octyl-agarose was added to 200 mL of lipase suspension 0.1 mg of protein mL at 48 C, pH 7.0 , in 10 mMphosphate and different concentrations of ammonium sulfate without ammonium sulfate , 0.1 M ammonium sulfate , 1 M ammonium sulfate A . Activity of supernatant was measured in all cases. 4. Periodically, withdraw samples using tip-cut pipets and measure the activity as described in Subheading...
Info Bto
BDNF, brain-derived neurotrofic factor BHK cells, baby hamster kidney cells BMP, bone morphogenetic protein EPO, erythropoietin GDNF, glial-cell derived neurotrofic factor MPS, mucopolysaccharidosis VE, vascular endothelium VEGF, vascular endothelial growth factor. BDNF, brain-derived neurotrofic factor BHK cells, baby hamster kidney cells BMP, bone morphogenetic protein EPO, erythropoietin GDNF, glial-cell derived neurotrofic factor MPS, mucopolysaccharidosis VE, vascular endothelium VEGF,...
Info Tns
Analysis, as described in the Subheading 3.1. is performed, and followed by addition of the ATP standard for internal calibration. After copmleting Subheading 3.1., step 7 add 10 L reconstituted ATP standard 10-6 M , mix quickly, place the cuvet back into the measurement chamber and measure light output. Increased response IR is given in RLU, which corresponds directly to the ATP level present in the sample and the ATP standard. ATP content in test is calculated by using RLU values of the...
Biomedical Applications of Immobilized Cells
Gorka Orive, Rosa Mar a Hern ndez, Alicia Rodr guez Gasc n, and Jos Luis Pedraz The aim of cell microencapsulation technology is to treat multiple diseases in the absence of immunosuppression. For this purpose, cells have been immobilized experimentally within carefully designed capsules that allow the long-term function of the graft. Recently, several advances have brought the whole technology much closer to a realistic proposal for clinical application. This chapter reviews the potential...
Xylanase Immobilization In 2000s
Fig. 1. Characterization of enzyme-polymer conjugate by A fluorescence spectroscopy difference spectrum of the immobilized and free enzymes and difference spectrum of the immobilized enzyme after subtracting the contribution of the polymer from the enzyme spectrum and B circular dichroic measurements. Xylanase was purified by affinity precipitation with Eudragit S-100, before being immobilized on Eudragit L-100. Equal amounts of protein were taken for I free as well as II immobilized enzyme....
Thermodynamic Equilibrium Constant Of Glutaraldehyde
then the thermodynamic parameters of the reaction e.g., free energy, enthalpy and entropy changes truly define the concept of stability. This definition is different from that used for kinetic experiments, where the biological activity is followed as a function of time in destabilizing conditions. In this case the reaction studied may be written as where I is the irreversibly inactivated form of the protein or enzyme. The time dependence of the activity gives information on the overall N I...
Immobilization of Cells on Polyurethane Foam
Ignacio de Ory, Gema Cabrera, Martin Ramirez, and Ana Blandino In this chapter, protocols and details for the immobilization of a model cell onto polyurethane foam carriers are provided in order to facilitate the use of such systems in laboratory or industrial reactors. Polyurethane foam has recently acquired great relevance as a carrier for its good mechanical properties, high porosity, and large adsorption surface. In addition, it has a very low commercial cost. Two different immobilization...
Info Wqh
Sepabead Histidine-tagged enzyme immobilization Multipoint covalent immobilization Ethanol, production with Lentikats-encapsulated yeast, 342 Ferredoxin-NADP reductase, gold support immobilization, 236 Fluidized bed reactors, advantages, 312, 314 examples of immobilized systems, 312, 313 magnetic particles, 314 pullulanase and glucoamylase mixed immobilization system, activity assays, 315 alginate bead entrapment, 315, 316 materials, 314-316 starch hydrolysis, 315-317 Formate dehydrogenase,...
Enzyme Immobilization Via Crosslinking Protocols
Fig. 4. Multipoint covalent attachment of proteins. the support, after which other subunits of each enzyme molecule can be cross-linked to the immobilized subunits via bifunctional, or polyfunctional, cross-linking agents e.g., glutaraldehyde, aldehyde-dextran . In this way, complex multimeric structures can be fully stabilized 4,5 see Fig. 5 . 4.3.2. CLEAs or CLECs of Multimeric Enzymes The aggregation or crystallization of multimeric enzymes with further cross-linking of the aggregated or...
Ab
Fig. 3. A SEM image of untreated SIRAN showing hollow pores in which the lipase-containing sol-gel material can be bound. B SEM image of a lipase SP 523 containing TMOS PDMS 4 1 gel in the pores of SIRAN. C Approximate 100-fold magnification of the SEM image of B 19 . Fig. 3. A SEM image of untreated SIRAN showing hollow pores in which the lipase-containing sol-gel material can be bound. B SEM image of a lipase SP 523 containing TMOS PDMS 4 1 gel in the pores of SIRAN. C Approximate 100-fold...
Info Tma
Acetate buffer 50 mM, pH 5.0. Flow and batch notations are referred to flow and batch calo-rimetry, respectively. Nis the number of the binding sites. From ref. 18. Copyright 2005, with permission from Elsevier. Acetate buffer 50 mM, pH 5.0. Flow and batch notations are referred to flow and batch calo-rimetry, respectively. Nis the number of the binding sites. From ref. 18. Copyright 2005, with permission from Elsevier. Inert supports can be used as protein and enzymes carriers for particular...
Purification Immobilization Hyperactivation and Stabilization of Lipases by
Jose M. Palomo, Gloria Fern ndez-Lorente, Cesar Mateo, Rosa L. Segura, Claudia Ortiz, Roberto Fernandez-Lafuente, and Jose M. Guisan Immobilization of lipases on hydrophobic supports at low ionic strength permits one-step purification, immobilization, hyperactivation, and stabilization of most lipases. This selective adsorption occurs because the hydrophobic surface of the supports is able to promote the interfacial activation of the lipases, yielding enzyme preparations having the open form of...
Characterization of Immobilized Enzymes by Microcalorimetry
Direct information on the stability and biological activity of immobilized proteins can be obtained from calorimetry. This technique is flexible enough to give insight on the thermodynamic consequences of the immobilization in most experimental conditions, ranging from multipoint covalent attachment to simple absorption. Different calorimetric techniques can be tailored to study the different aspects of the protein chemistry, depending on the physical environment and the type of confinement....
AgAgCI
Fig. 2. Two-channel screen-printed electrodes used for AChE-biosensor construction. and enzyme activity may be determined with these solutions by employing the Ellman method 9 . 3.1. Preparation of the Screen-Printed Electrodes A single-channel two-electrode sensing strip 7 mm x 44 mm x 0.5 mm will be used in this work. Figure 2 presents a two-channel sensor strip, prepared as follows 10,11 1. Use PVC supports to prepare the strips with a DEK 248 screen-printer DEK . 2. Print the conducting...