Endo‐β‐Glucosidase Tag Allows Dual Detection of Fusion Proteins by Fluorescent Mechanism‐Based Probes and Activity Measurement

β‐Glucoside‐configured cyclophellitols are activity‐based probes (ABPs) that allow sensitive detection of β‐glucosidases. Their applicability to detect proteins fused with β‐glucosidase was investigated in the cellular context. The tag was Rhodococcus sp. M‐777 endoglycoceramidase II (EGCaseII), based on its lack of glycans and ability to hydrolyze fluorogenic 4‐methylumbelliferyl β‐d ‐lactoside (an activity absent in mammalian cells). Specific dual detection of fusion proteins was possible in vitro and in situ by using fluorescent ABPs and a fluorogenic substrate. Pre‐blocking with conduritol β‐epoxide (a poor inhibitor of EGCaseII) eliminated ABP labeling of endogenous β‐glucosidases. ABPs equipped with biotin allowed convenient purification of the fusion proteins. Diversification of ABPs (distinct fluorophores, fluorogenic high‐resolution detection moieties) should assist further research in living cells and organisms.     

Effects of plasma treatment on biocompatibility of poly[(L‐lactide)‐co‐(ϵ‐caprolactone)] and poly[(L‐lactide)‐co‐glycolide] electrospun nanofibrous membranes

Poly[(l ‐lactide)‐co ‐(? ‐caprolactone)] (PLCL) and poly[(l ‐lactide)‐co ‐glycolide] (PLGA) copolymers are widely used in neural guide tissue regeneration. In this research, the surface modification of their hydrophilicity was achieved using plasma treatment. Attachment and proliferation of olfactory ensheathing cells on treated electrospun membranes increased by 26 and 32%, respectively, compared to the untreated PLCL and PLGA counterparts. Cells cultivated on both the PLCL and PLGA membranes showed high viability (>95%) and healthy morphologies with no evidence of cytotoxic effects. Cells grown on treated electrospun fibres displayed significant increases in mitochondrial activity and reductions in membrane leakage when compared to untreated samples. The results suggested that plasma treatment of the surface of the polymers enhanced both cell viability and growth without incurring any cytotoxic effects. © 2017 Society of Chemical Industry     

Synthesis of zwitterionic stationary phase based on hydrophilic non‐porous poly(glycidymethacrylate‐co‐ethylenedimethacrylate) beads and their application for fast separation of proteins

A new hydrophilic strong/strong type zwitterionic stationary phase for high performance liquid chromatography (HPLC) was synthesized by chemical modification of 3.0 μm non‐porous monodisperse poly(glycidylmethacrylate‐co‐ethylenedimethacrylate)(P_GMA/EDMA) beads in the following steps. First, the beads were reacted with hydrochloride to obtain chlorizated beads; second, chlorizated beads were reacted with dimethylamine to obtain ammoniated beads; third, ammoniated beads were reacted with 1,3‐propanesultone to obtain non‐porous hydrophilic zwitterionic stationary phase. The stationary phase was evaluated in detail to determine its ion‐exchange properties, separability, reproducibility, hydrophilicity, and the effect of column loading and pH on the separation and retention of proteins. The highest dynamic protein loading capacity of the synthesized zwitterionic packing for bovin serum albumin and Lys were 18.3 and 27.4 mg g^?1, respectively. The zwitterionic stationary phase was capable of separating two acidic and three basic proteins simultaneously in less than 2.5 min by the flow‐rates of 3.0 mL min^?1. The zwitterionic resin was also used for rapid separation and purification of recombinant human interferon‐r (rhIFN‐r) and human granulocyte colony‐stimulation factor (hG‐CSF) from the crude extract solution. The satisfactory results were obtained. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

N‐Lauroylation during the Expression of Recombinant N‐Myristoylated Proteins: Implications and Solutions

Incorporation of myristic acid onto the N terminus of a protein is a crucial modification that promotes membrane binding and correct localization of important components of signaling pathways. Recombinant expression of N‐myristoylated proteins in Escherichia coli can be achieved by co‐expressing yeast N‐myristoyltransferase and supplementing the growth medium with myristic acid. However, undesired incorporation of the 12‐carbon fatty acid lauric acid can also occur (leading to heterogeneous samples), especially when the available carbon sources are scarce, as it is the case in minimal medium for the expression of isotopically enriched samples. By applying this method to the brain acid soluble protein 1 and the 1–185 N‐terminal region of c‐Src, we show the significant, and protein‐specific, differences in the membrane binding properties of lauroylated and myristoylated forms. We also present a robust strategy for obtaining lauryl‐free samples of myristoylated proteins in both rich and minimal media.     

Design Strategies for Integrated β‐Galactosidase Purification Processes

The operational conditions for an aqueous two‐phase system (ATPS) for β‐galactosidase purification were optimized and applied to the design of a purification strategy as an alternative to the primary purification steps. The ATPS proved to be suitable for the recovery and primary enzyme purification. The purification process design developed by ATPS, diafiltration, ion exchange, and diafiltration/ultrafiltration was successful, yielding a more than tenfold purification. The purification strategy design resulted in a powerful integrated purification and recovery process, an evidence of the potential for a scale‐up of the β‐galactosidase purification process.     

Study of the effect of organic clay on the shear‐induced crystallization of high‐density polyethylene through dynamic‐packing injection molding

Organic nanoparticles as heterogeneous nucleators have a great effect on the crystallization of polymer matrices in nanocomposite systems, and the effect will be enhanced under shear flow. A home‐made dynamic‐packing injection molding (DPIM) device was developed to explore the effect of organic clay on the shear‐induced crystallization of high‐density polyethylene (HDPE). Differential scanning calorimetry (DSC), wide‐angle X‐ray diffraction (WAXD) and scanning electron microscopy (SEM) were used to characterize the flow‐induced crystalline structure of HDPE/clay nanocomposite injection moldings. It was found that higher crystallinity and thicker crystal planes which contribute to the improvement of mechanical properties were achieved in HDPE/clay nanocomposite samples prepared by DPIM. DSC results clearly showed that an increase of about 16% in crystallinity was achieved in dynamic HDPE/clay nanocomposite samples compared with traditional unfilled HDPE samples. WAXD confirmed that dynamic HDPE/clay nanocomposite samples had maximum crystal sizes at the (110) and (200) planes of 335 and 305 Å, respectively. SEM images indicated that the arrangement of crystalline structures in dynamic HDPE/clay samples was altered slightly compared with unfilled HDPE samples prepared using the same processing parameters. The results showed that organic clay was beneficial for increasing crystallinity and crystal size in the HDPE/clay nanocomposite system under shear flow. Meanwhile the arrangement of crystalline structures was insignificantly affected by the organic clay, and the preferred regular arrangement of lamellae could still be formed in the dynamic HDPE/clay nanocomposite system. Copyright © 2010 Society of Chemical Industry     

Siloxane‐modified polyacrylate low‐residual pressure‐sensitive adhesive with high peeling strength

A low‐residual siloxane‐modified polyacrylate pressure‐sensitive adhesive (PSA) with a high peeling strength was prepared by seeded semicontinuous emulsion polymerization. 3‐Glycidyloxypropyltrimethoxysilane was introduced into the acrylic (AC) PSA through a thermal posttreatment method to crosslink with AC. To improve the adhesion properties, a polymeric emulsifier, 2‐acrylamido‐2‐methyl‐1‐propanesulfonic acid, was introduced into the system with the conventional emulsifier. Several key polymerization conditions, such as the initiator concentration, mass ratio of soft monomer to hard monomer, the content of polymeric emulsifier, and siloxane dosages were examined in detail. Then, the optimal conditions and a proper preparation process were established. The results show that we achieved not only a low repeeling residue with high tack and peeling strength but also excellent properties of high‐temperature aging resistance and water resistance. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 42975.     

Synthesis,characterization and application of poly[(1‐vinyl‐2‐pyrrolidone)‐co‐(2‐hydroxyethyl methacrylate)] as controlled‐release polymeric system for 2,4‐dichlorophenoxyacetic chloride using an ultrafiltration technique

BACKGROUND: Polymers supporting chemicals used in agriculture have recently been developed to overcome the serious environmental problems of conventional agrochemicals. The success of these formulations is based on a suitable choice of polymer support. Degradable polymeric hydrogels are of particular interest. The gradual release of the bioactive agent can be achieved by hydrolytic or enzymatic cleavage of the linking bond. RESULTS: In this context, poly[(1‐vinyl‐2‐pyrrolidone)‐co‐(2‐hydroxyethyl methacrylate)] [poly(NVP‐co‐HEMA)] has been used as a bioactive carrier reagent. Herein, we report a controlled‐release system with the herbicide 2,4‐dichlorophenoxyacetic acid (2,4‐D) using an ultrafiltration system. Hydrolysis was studied by testing the release at various pH values. A high release with poly(NVP‐co‐HEMA)–2,4‐D was observed at pH = 7 and 10 after two days (Z = 2). The release percentage of copolymer–herbicide increased at pH = 10. It showed release values between 79.0 and 94.5%. Poly(NVP‐co‐HEMA)–herbicide can release a bioactive compound in aqueous solution at pH = 3, 7 and 10. CONCLUSION: Based on the results of homogeneous hydrolysis, it is argued that the herbicide release rate depends on the pH of the reaction environment. This functional polymer could be employed as a biodegradable material for applications in agrichemical release. Copyright © 2008 Society of Chemical Industry     

Efficient mAb production in CHO cells incorporating PEI‐mediated transfection,mild hypothermia and the co‐expression of XBP‐1

BACKGROUND: Transient gene expression (TGE) provides a rapid way to generate recombinant protein biologics for pre‐clinical assessment. Human embryonic kidney (HEK293) cells have traditionally been used for TGE; however, there is demand from industry for efficient, high‐producing TGE systems that utilize Chinese hamster ovary (CHO) cells. A polyethyleneimine (PEI) ‐based TGE process has been developed for CHO cells using an episomal expression system to generate enhanced recombinant protein titers. RESULTS: A five‐fold improvement in monoclonal antibody (mAb) volumetric productivity was achieved by examining key parameters including transfection medium, cell density, transfection reagent, DNA:reagent ratio, the time of transfer to mild hypothermia and feeding strategy post‐transfection. The Epi‐CHO system allowed for a six‐fold expansion in culture volume post‐transfection without significantly affecting specific productivity. This system generates 400% more mAb per µg of plasmid DNA when compared with a non‐episomal system. In addition, the use of X‐box binding protein 1 to enhance secretion capacity and provide further improvements in mAb production with TGE was investigated. CONCLUSION: Through optimization of key parameters, our results demonstrate the development of a low‐cost, high‐yielding, episomal TGE system that may be adopted during pre‐clinical biologic drug development. Copyright © 2011 Society of Chemical Industry     

Simultaneous refolding,purification, and immobilization of recombinant Fibrobacter succinogenes 1,3‐1,4‐β‐D‐glucanase on artificial oil bodies

BACKGROUND: 1,3‐1,4‐β‐D‐glucanase (1,3‐1,4‐β‐D‐glucan 4‐glucanohydrolase; EC 3.2.1.73) has been used in a range of industrial processes. As a biocatalyst, it is better to use immobilized enzymes than free enzymes, therefore, the immobilization of 1,3‐1,4‐β‐D‐glucanase was investigated. RESULTS: A 1,3‐1,4‐β‐D‐glucanase gene from Fibrobacter succinogenes was overexpressed in Escherichia coli as a recombinant protein fused to the N terminus of oleosin, a unique structural protein of seed oil bodies. With the reconstitution of the artificial oil bodies (AOBs), refolding, purification, and immobilization of active 1,3‐1,4‐β‐D‐glucanase was accomplished simultaneously. Response surface modeling (RSM), with central composite design (CCD), and regression analysis were successfully applied to determine the optimal temperature and pH conditions of the AOB‐immobilized 1,3‐1,4‐β‐D‐glucanase. The optimal conditions for the highest immobilized 1,3‐1,4‐β‐D‐glucanase activity (7.1 IU mg^?1 of total protein) were observed at 39 °C and pH 8.8. Furthermore, AOB‐immobilized 1,3‐1,4‐β‐D‐glucanase retained more than 70% of its initial activity after 120 min at 39 °C, and it was easily and simply recovered from the surface of the solution by brief centrifugation; it could be reused eight times while retaining more than 80% of its activity. CONCLUSIONS: These results indicate that the AOB‐based system is a comparatively simple and effective method for simultaneous refolding, purification, and immobilization of 1,3‐1,4‐β‐D‐glucanase. Copyright © 2009 Society of Chemical Industry     

Development of a pre‐purification process for homoharringtonine

A novel pre‐purification method was developed for producing homoharringtonine from Cephalotaxus koreana, giving high purity and yield. The simple, efficient procedure involved biomass extraction, liquid–liquid extraction, and synthetic adsorbent treatment, followed by low‐pressure chromatography. The use of active clay treatment and silica gel low‐pressure chromatography in the pre‐purification process allowed for the rapid, efficient separation of homoharringtonine from interfering compounds and, compared with alternative processes, increased the yield and purity of crude homoharringtonine for subsequent high‐performance liquid chromatography (HPLC) purification. Homoharringtonine of over 52% purity could be obtained simply with high yield from biomass using this pre‐purification method, while minimizing solvent use and the scale and complexity of HPLC operations for homoharringtonine purification. Copyright © 2005 Society of Chemical Industry     

Large‐scale isolation and purification of C‐phycocyanin from the cyanobacteria Anabaena marina using expanded bed adsorption chromatography

BACKGROUND: Phycobiliproteins are water soluble proteins useful as fluorescent markers of cells and macromolecules, and as natural colorants, and are anticarcinogenic. Although phycobiliproteins have many applications, their use is limited by the high cost of the purified macromolecules, mainly related with the cost of extraction and purification. In this study a fast and scalable method for preparative extraction and purification of C‐phycocyanin (C‐PC) from Anabaena marina is developed. RESULTS: The method developed consists in the extraction of phycobiliproteins using repeated single contact strategy, separation being performed by expanded bed adsorption (EBA) chromatography using Streamline‐DEAE. Optimal conditions for EBA were obtained at small scale, using a 15 mm internal diameter column, these being a sample load of 0.9 mg C‐PC mL^?1 adsorbent, an expanded bed volume twice the settled bed volume and a sample viscosity of 1.109 mP. The process was then scaled up 36 times, the success of the scale‐up process being verified. Finally, to obtain pure C‐PC conventional ion‐exchange chromatography was utilized. CONCLUSION: Small diameter columns was shown to be useful to simulate the behavior of larger diameter columns for use in scaled up systems. Expanded bed adsorption was demonstrated to be a scalable technology allowing large quantities of C‐PC to be obtained, maintaining high protein recovery while reducing both processing cost and time. The proposed methodology allows recovery of more than 62% of the C‐PC contained in the biomass in the form of pure C‐PC concentrates. Copyright © 2010 Society of Chemical Industry     

Irradiation grafting of methyl methacrylate monomer onto ultra‐high‐molecular‐weight polyethylene: An experimental design approach for improving adhesion to bone cement

The grafting of methyl methacrylate (MMA) onto ultra‐high‐molecular‐weight polyethylene (UHMWPE) and chromic acid etched UHMWPE was conducted with a preirradiation method in air in the presence of a Mohr salt and sulfuric acid. The grafted samples were characterized by Fourier transform infrared (FTIR) spectroscopy, a gravimetric method, differential scanning calorimetry, scanning electron microscopy (SEM), and interfacial bonding strength measurements. The FTIR results showed the presence of ether and carbonyl groups in the MMA‐grafted UHMWPE (MMA‐g‐UHMWPE) samples. The Taguchi experimental design method was used to find the best degree of grafting (DG) and bonding strength. The efficient levels for different variables were calculated with an analysis of variance of the results. SEM micrographs of MMA‐g‐UHMWPE samples showed that with increasing DG and chromic acid etching, the MMA‐g‐UHMWPE rich phase increased on the surface; this confirmed the high interfacial bonding strength of the grafted samples with bone cement. The grafting of the MMA units onto UHMWPE resulted in a lower crystallinity, and the crystallization process proceeded at a higher rate for the MMA‐g‐UHMWPE samples compared to the initial UHMWPE; this suggested that the MMA grafted units acted as nucleating agents for the crystallization of UHMWPE. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Two‐Way Reversible Shape Memory Properties of Benzoyl Peroxide Crosslinked Poly(ethylene‐co‐vinyl acetate) under Different Stress Conditions

Two‐way (reversible) chemically crosslinked semicrystalline shape memory polymers are synthesized using poly(ethylene‐co‐vinyl acetate) (PEVA) with benzoyl peroxide (BPO). The two‐way shape memory effect (2W‐SME) is achieved under both constant stress and stress‐free conditions. It is found that the stress‐free 2W‐SME can be achieved by the relationship between the initial prestretching strain (R_prestretch) and recovery strain (R_rec). Under the same prestretching stress, the stress‐free two‐way shape memory behavior can be controlled by variation of R_rec using a different setting temperature (T_set) in the recovery process. More importantly, the driving force and recovery force, as one of the key indicators for two‐way shape memory materials, are investigated, and they significantly change depending on the BPO content. The sample with high BPO content shows excellent high‐temperature creep resistant performance. A highly crosslinked structure can suppress viscous flow and provides sufficient force to allow the sample to recover its initial shape after crystal melting. Therefore, the PEVA/BPO samples are able to contract during heating. The presence of an oriented crystal structure with high applied stress that causes sample elongation during cooling is also investigated. These findings for PEVA/BPO two‐way shape memory polymers will contribute to their applications as soft actuators in various fields.     

Graft copolymers and high‐molecular‐weight star‐like polymers by atom transfer radical polymerization

Grafting of tert‐butyl acrylate (tBuA), methyl methacrylate (MMA), and styrene (St) monomers (M) by Cu(I)‐mediated ATRP from polystyrene (PSt) macroinitiator (M_n = 5620, polydispersity index, PDI = 1.12), containing initiating 2‐bromopropionyloxy groups (I) (bound to 34% of aromatic cores; 11 groups per backbone), was performed using conditions suitable for the respective homopolymerizations. The preparation of PSt‐g‐PtBuA in bulk using an initial molar ratio [M]₀/[I]₀ = 140 had a controlled character up to M_n = (132–148) × 10³ (PDI = 1.08–1.16). With MMA and St and using the same [M]₀/[I]₀, preliminary experiments were made; the higher the monomer conversion, the broader was the distribution of molecular weight of the products. Graft copolymerizations of all these monomers at [M]₀/[I]₀ = 840 or 1680 were successfully conducted up to high conversions. Low‐polydispersity copolymers, with very long side chains, in fact star‐like copolymers, were obtained mainly by tuning the deactivator amount in the reaction mixture. (PSt‐g‐PtBuA, DP_n,sc (DP of side chain) = 665, PDI = 1.24; PSt‐g‐PMMA, DP_n,sc = 670, PDI = 1.43; PSt‐g‐PSt, DP_n,sc = 324, PDI = 1.11). Total suppression of intermolecular coupling was achieved here. However, the low concentrations of initiator required long reaction times, leading sometimes to formation of a small amount (～5%) of low‐molecular‐weight polymer fraction. This concomitant process is discussed, and some measures for its prevention are proposed. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 3662–3672, 2006     

Iron‐Catalyzed Oxidative Cleavage of Olefins and Alkynes to Carboxylic Acids with Aqueous tert‐Butyl Hydroperoxide

A new method for the oxidation of aromatic olefins and alkynes has been developed using inexpensive iron(III) chloride hexahydrate (FeCl₃⋅6 H₂O, 5 mol%) catalyst in combination with commercially available aqueous 70% tert‐butyl hydroperoxide (TBHP) as oxidant. The present system works well for aromatic alkenes and alkynes with both electron‐donating and electron‐withdrawing substituents being tolerated. The protocol is free from chromatographic purification and the carboxylic acids are obtained in high yields by simple filtration.     

Cationic Gold‐ and Silver‐Catalyzed Cycloisomerizations of Propargylic Ureas: A Selective Entry to Oxazolidin‐2‐imines and Imidazolidin‐2‐ones

A comparative study on transition metal‐catalyzed cycloisomerizations of propargylic ureas derived in situ from secondary propargylic amines and tosyl isocyanate was performed. The influence of catalytic system on the reaction outcome was thoroughly studied on two model examples resulting in the establishment of two selective protocols for both O‐ and N‐cyclizations. The application of cationic gold(I) catalysis generally resulted in a formation of oxazolidin‐2‐imines as major products while the application of silver(I) triflate selectively provided the corresponding imidazolidin‐2‐ones. An attempt to rationalize the observed chemoselectivity is described. The scope of both processes was demonstrated through the use of variously substituted secondary propargylic amines.     

Polymer‐Supported,Carbon Dioxide‐Protected N‐Heterocyclic Carbenes: Synthesis and Application in Organo‐ and Organometallic Catalysis

The synthesis of a resin‐supported, carbon dioxide‐protected N‐heterocyclic carbene (NHC) and its use in organocatalysis and organometallic catalysis are described. The resin‐bound carbon dioxide‐protected NHC‐based catalyst was prepared via ring‐opening metathesis copolymerization of 1,4,4a,5,8,8a‐hexahydro‐1,4,5,8‐exo,endo‐dimethanonaphthalene ( DMNH6 ) with 3‐(bicyclo[2.2.1]hept‐5‐en‐2‐ylmethyl)‐1‐(2‐propyl)‐3,4,5,6‐tetrahydropyrimidin‐1‐ium‐2‐carboxylate ( M1 ), using the well‐defined Schrock catalyst Mo[N‐2,6‐(2‐Pr)₂‐C₆H₃](CHCMe₂Ph)(OCMe₃)₂ and was used for a series of organocatalytic reactions, i.e., for the trimerization reaction of isocyanates, as well as for the cyanosilylation of carbonyl compounds. In the latter reaction, turn‐over numbers (TON) up to 5000 were achieved. In addition, the polymer‐supported, carbon dioxide‐protected N‐heterocyclic carbene served as an excellent progenitor for various polymer‐supported metal complexes. It was loaded with a series of rhodium(I), iridium(I), and palladium(II) precursors and the resulting Rh‐, Ir‐, and Pd‐loaded resins were successfully used in the polymerization of phenylacetylene, in the hydrogen transfer reaction to benzaldehyde, as well as in Heck‐type coupling reactions. In the latter reaction, TONs up to 100,000 were achieved. M1 , as a non‐supported analogue of poly‐M1‐b‐DMNH6 , as well as the complexes PdCl₂[1,3‐bis(2‐Pr)tetrahydropyrimidin‐2‐ylidene]₂ ( Pd‐1 ) and IrBr[1‐(norborn‐5‐ene‐2‐ylmethyl)‐3‐(2‐Pr)‐3,4,5,6‐tetrahydropyrimidin‐2‐ylidine](COD) ( Ir‐1 ) were used as homogeneous analogues and their reactivity in the above‐mentioned reactions was compared with that of the supported catalytic systems. In all reactions investigated, the TONs achieved with the supported systems were very similar to the ones obtained with the unsupported, homogeneous ones, the turn‐over frequencies (TOFs), however, were lower by up to a factor of three.     

Gold(I)‐Catalyzed Regio‐ and Stereoselective 1,3‐Dipolar Cycloaddition Reactions of 1‐(1‐Alkynyl)cyclopropyl Oximes with Nitrones: A Modular Entry to Highly Substituted Pyrrolo[3,4‐d][1,2]oxazepines

An efficient approach for the synthesis of highly substituted pyrrolo[3,4‐d][1,2]oxazepines has been achieved by gold(I)‐catalyzed 1,3‐dipolar cycloaddition reactions of 1‐(1‐alkynyl)cyclopropyl oximes with nitrones in good to excellent yields as a single diastereomer. A complete chirality transfer was observed in this transformation.     

Adsorption of blood proteins on glow‐discharge‐modified polyurethane membranes

Polyurethanes are a class of polymers that have a wide range of applications in the medical field although their blood compatibility still needs improvement. In order to obtain medical purity, this study prepared membrane‐form polyurethanes from toluene 2,4‐diisocyanate (TDI) and poly(propylene ethylene glycol) without the addition of any ingredients such as solvents, catalysts, or chain extenders. The aim was to increase surface hydrophilicity and improve blood compatibility. Therefore, the prepared membranes were modified by treatment with oxygen or argon plasmas. Characterizations of the samples were achieved by contact‐angle and water‐uptake studies as well as from atomic force microscope (AFM) pictures. It was found that oxygen‐modified samples were more hydrophilic than argon‐modified samples. The AFM images showed that surface roughness increased with plasma treatment. The protein adsorption experiments carried out with single protein solutions demonstrated that the adsorption of bovine serum albumin and fibrinogen decreased drastically by increasing the applied power and exposure time of the glow discharge. A similar decrease in the adsorption of protein was also observed for human blood proteins. The alterations of the conformational structures of the adsorbed proteins were examined by fluorescence spectrophotometry. Similar spectra with the same maximum wavelength were observed for native and desorbed proteins. These results showed that no denaturation of the proteins occurred upon adsorption on the surfaces of the prepared membranes. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1322–1332, 2001