The inactivation of horseradish peroxidase by a polystyrene surface

Berkowitz, David B.; Webert, Donald W.

doi:10.1016/0022-1759(81)90263-5

Cited by 22 publications

(6 citation statements)

References 4 publications

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“…When the solution interactions are stronger, the protein remains in solution. There are some models of protein attachment where the protein molecule is considered as rigid. − However, the protein molecule is flexible and the conformation depends on environment (solution, pH, ion strength, and so on), temperature, and presence of specific molecules on the surface. − Therefore, surface immobilization often affects protein conformation and activity.…”

Section: Discussionmentioning

confidence: 99%

Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

2011

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Horseradish peroxidase (HRP) was immobilized onto both plasticized and unplasticized polyvinylchloride (PVC) and ultrahigh molecular weight polyethylene (UHMWPE). Plasma immersion ion implantation (PIII) in a nitrogen plasma with 20 kV bias was used to facilitate covalent immobilization and to improve the wettability of the surfaces. The surfaces and immobilized protein were studied using attenuated total reflection infrared (ATR-IR) spectroscopy and water contact angle measurements. Protein elution on exposure to repeated sodium dodecyl sulfate (SDS) washing was used to assess the strength of HRP immobilization. The presence of low molecular weight components (plasticizer, additives in solvent, unreacted monomers, adsorbed molecules on surface) was found to have a major influence on the strength of immobilization and the conformation of the protein on the samples not exposed to the PIII treatment. A phenomenological model considering interactions between the low molecular weight components, the protein molecule, and the surface is developed to explain these observations.

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Section: Discussionmentioning

confidence: 99%

Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

2011

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“…Knowledge of the conformation of an attached protein is important to understand the activity of the protein molecule and its functionality in a biological system. Attached HRP protein on untreated hydrocarbon polymeric materials does not preserve its native conformation [94]. We have investigated the conformation of attached protein on PIII treated polyethylene using the Amide I line in FTIR ATR spectroscopy.…”

Section: Conformation Of Attached Proteinmentioning

confidence: 99%

Mechanisms for Covalent Immobilization of Horseradish Peroxidase on Ion-Beam-Treated Polyethylene

et al. 2012

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The surface of polyethylene was modified by plasma immersion ion implantation. Structure changes including carbonization and oxidation were observed. High surface energy of the modified polyethylene was attributed to the presence of free radicals on the surface. The surface energy decay with storage time after treatment was explained by a decay of the free radical concentration while the concentration of oxygen-containing groups increased with storage time. Horseradish peroxidase was covalently attached onto the modified surface by the reaction with free radicals. Appropriate blocking agents can block this reaction. All aminoacid residues can take part in the covalent attachment process, providing a universal mechanism of attachment for all proteins. The native conformation of attached protein is retained due to hydrophilic interactions in the interface region. The enzymatic activity of covalently attached protein remained high. The long-term activity of the modified layer to attach protein is explained by stabilisation of unpaired electrons in sp2 carbon structures. A high concentration of free radicals can give multiple covalent bonds to the protein molecule and destroy the native conformation and with it the catalytic activity. The universal mechanism of protein attachment to free radicals could be extended to various methods of radiation damage of polymers.

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“…Altered recognition of solid-phase antigens has been reported (Dierks et al, 1986). The inclusion of the anionic detergent Tween-20 appears to stabilize HRP conjugates when adsorbed onto plastic (Berkowitz and Webert, 1981) and may be essential to obtain consistent results. In the assays described in this paper, we included Tween-20 in buffers used for washing plates and for diluting samples.…”

Section: Discussionmentioning

confidence: 99%

Definition of functional and antibody-binding sites on Kunitz soybean trypsin inhibitor isoforms using monoclonal antibodies

Brandon¹,

Bates²

1988

J. Agric. Food Chem.

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The inactivation of horseradish peroxidase by a polystyrene surface

Cited by 22 publications

References 4 publications

Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

Mechanisms for Covalent Immobilization of Horseradish Peroxidase on Ion-Beam-Treated Polyethylene

Definition of functional and antibody-binding sites on Kunitz soybean trypsin inhibitor isoforms using monoclonal antibodies

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