We report on the successful application of carboxyl-rich plasma polymerized (PP) films as a matrix layer for bioreceptor immobilization in surface plasmon resonance (SPR) immunosensing. Composition and chemical properties of the carboxyl-rich PP films deposited from a mixture of maleic anhydride and acetylene were investigated. Changes in the films stored in air, water, and buffer were studied and the involved chemical changes were described. Performance in SPR immunosensing was evaluated on interactions of human serum albumin (HSA) with a specific monoclonal antibody. The comparison with the mixed self-assembled monolayer of mercaptoundecanoic acid and mercaptohexanol (MUA/MCH) and one of the most widely used surfaces for SPR, the 2D and 3D carboxymethylated dextran (CMD), was presented to show the efficacy of plasma polymerized matrix layers for biosensing. The PP film-based SPR immunosensor provided a similar detection limit of HSA (100 ng/mL) as MUA/MCH- (100 ng/mL) and 3D CMD (50 ng/mL)-based sensors. However, the response levels were about twice higher in case of the PP film-based immunosensor than in case of MUA/MCH-based alternative. The PP film surfaces had similar binding capacity towards antibody as the 3D CMD layers. The response of PP film-based sensor towards HSA was comparable to 3D CMD-based sensor up to 2.5 μg/mL. For the higher concentrations (> 10 μg/mL), the response of PP film-based immunosensor was lower due to inaccessibility of active sites of the immobilized antibody inside the flat PP film surface. We have demonstrated that due to its high stability and cost-effective straightforward preparation, the carboxyl-rich PP films represent an efficient alternative to self-assembled monolayers (SAM) and dextran-based layers in label-free immunosensing. Graphical abstract.
- MeSH
- Acetylene chemistry MeSH
- Biosensing Techniques MeSH
- Limit of Detection MeSH
- Maleic Anhydrides chemistry MeSH
- Microscopy, Atomic Force MeSH
- Plasma Gases * MeSH
- Polymers chemistry MeSH
- Surface Plasmon Resonance methods MeSH
- Surface Properties MeSH
- Sulfhydryl Compounds chemistry MeSH
- Publication type
- Journal Article MeSH
We introduce a stable isotope labeling approach for glycopeptides that allows a specific glycosylation site in a protein to be quantitatively evaluated using mass spectrometry. Succinic anhydride is used to specifically label primary amino groups of the peptide portion of the glycopeptides. The heavy form (D4(13)C4) provides an 8 Da mass increment over the light natural form (H4(12)C4), allowing simultaneous analysis and direct comparison of two glycopeptide profiles in a single MS scan. We have optimized a protocol for an in-solution trypsin digestion, a one-pot labeling procedure, and a post-labeling solid-phase extraction to obtain purified and labeled glycopeptides. We provide the first demonstration of this approach by comparing IgG1 Fc glycopeptides from polyclonal IgG samples with respect to their galactosylation and sialylation patterns using MALDI MS and LC-ESI-MS.
- MeSH
- Succinic Anhydrides chemistry MeSH
- Glycopeptides chemistry MeSH
- Glycosylation MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Immunoglobulin G chemistry MeSH
- Isotope Labeling MeSH
- Humans MeSH
- Protein Processing, Post-Translational * MeSH
- Proteomics methods MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Esterifications of the tree-based gum, gum karaya (GK), using dodecenylsuccinic anhydride (DDSA) were carried out in aqueous solutions. GK was deacetylated using alkali treatment to obtain deacetylated gum karaya (DGK). The DGK and its DDSA derivative were characterized using gel permeation chromatography/multiangle laser light scattering (GPC/MALLS), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), proton nuclear magnetic resonance spectroscopy ((1)H NMR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) analysis, and rheological studies. The degree of substitution was found to be 10.25% for DGK using (1)H NMR spectroscopy. The critical aggregation concentration of DDSA-DGK was determined using dye solubilization and surface tension methods. The antibacterial activity of the DDSA-DGK derivative was then investigated against Gram-negative Escherichia coli and Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus. The DDSA-DGK derivative has the potential for use as a stabilizing agent in food and nonfood applications. It can also be developed as an antibacterial agent.
- MeSH
- Succinic Anhydrides chemistry MeSH
- Anti-Bacterial Agents chemistry isolation & purification pharmacology MeSH
- Bacteria drug effects MeSH
- Karaya Gum chemistry isolation & purification pharmacology MeSH
- Plant Extracts chemistry isolation & purification pharmacology MeSH
- Sterculia chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Polymer therapeutics including polymer-drug conjugates, polymer-protein conjugates and polymer-modified gene delivery vectors are addressed in this review. Brief history of the polymer therapeutics is described with a focus on the pioneering work accomplished at the Institute of Macromolecular Chemistry in Prague. The advantages of polymer therapeutics compared with lowmolecular- weight drugs are outlined. Polymer cancerostatics, polymer-protein conjugates with anti-cancer activity and polymer-modified viruses based on N-(2-hydroxypropyl) methacrylamide copolymers and biodegradable multiblock poly(ethylene glycol) polymers are chosen as examples. The current status of clinical evaluation of the polymer therapeutics is also mentioned.
- MeSH
- Biopolymers chemistry classification therapeutic use MeSH
- Cytostatic Agents MeSH
- Ethylene Glycols chemistry therapeutic use MeSH
- Financing, Organized MeSH
- Genetic Therapy methods MeSH
- Pharmaceutical Preparations MeSH
- Humans MeSH
- Maleic Anhydrides therapeutic use MeSH
- Methacrylates chemistry therapeutic use MeSH
- Polymers history chemistry therapeutic use MeSH
- Polystyrenes therapeutic use MeSH
- Zinostatin therapeutic use MeSH
- Check Tag
- Humans MeSH
Concise international chemical assessment document ; 75
iv, 60 s. : il. ; 30 cm
- MeSH
- Anhydrides adverse effects toxicity MeSH
- Risk Assessment MeSH
- Maximum Allowable Concentration MeSH
- Environmental Exposure MeSH
- Conspectus
- Životní prostředí a jeho ochrana
- NML Fields
- environmentální vědy
- NML Publication type
- publikace WHO
