5 results
Search Results
2. Instant on-paper protein digestion during blood spot sampling.
- Author
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Skjærvø Ø, Rosting C, Halvorsen TG, and Reubsaet L
- Subjects
- Amino Acid Sequence, Humans, Paper, Peptides chemistry, Trypsin, Dried Blood Spot Testing, Proteins chemistry, Proteolysis
- Abstract
A concept integrating sampling and protein digestion is introduced here combining fast and simple fabrication by wax printing on filter paper with trypsin immobilized polymer beads. The paper reactors showed promising results with a high degree of protein digestion within fifty minutes in model protein mixtures as well as in human blood. The model protein mixture was used for the evaluation of performance both with and without a reduction and alkylation step. The paper reactors without reduction and alkylation showed between 46% and 75% protein sequence coverage and between five and 20 high confidence peptides (one and five zero missed cleavage peptides, respectively). Compared to a conventional in-solution approach, the paper reactor showed 10% less protein sequence coverage, 29% fewer high confidence peptides and 19% fewer high confidence peptides with zero missed cleavages. Placement of the protein reduction and alkylation step (before or after protein digestion) was shown to be of low importance. The storage stability of the paper reactors with (six weeks) and without (twelve weeks) tryptic peptides was satisfactory. The ability of the paper reactors to digest complex biological samples was investigated by comparison with human whole blood samples prepared using a conventional dried blood spot (DBS) procedure with overnight digestion in non-targeted analysis. The reactors showed a comparable performance with 75 ± 25 for the protein groups compared to 76 ± 5 for the DBS samples. Additionally, 267 ± 72 and 335 ± 11 unique peptides (high confidence) were identified for on-paper digestion and DBS, respectively.
- Published
- 2017
- Full Text
- View/download PDF
3. Laboratory-scale protein striping system for patterning biomolecules onto paper-based immunochromatographic test strips.
- Author
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Nash MA, Hoffman JM, Stevens DY, Hoffman AS, Stayton PS, and Yager P
- Subjects
- Animals, Antibodies, Immobilized chemistry, Antibodies, Immobilized immunology, Biosensing Techniques economics, Clinical Laboratory Techniques, Equipment Design, Gold chemistry, Humans, Immunoassay economics, Immunoglobulin G chemistry, Immunoglobulin G immunology, Metal Nanoparticles chemistry, Proteins immunology, Streptavidin chemistry, Streptavidin immunology, Syringes, Time Factors, Biosensing Techniques instrumentation, Immunoassay instrumentation, Paper, Proteins chemistry, Reagent Strips
- Abstract
A method for patterning narrow lines of biomolecules onto nitrocellulose membranes using laboratory syringe pumps is described. One syringe pump is used to drive the biomolecule solution through a needle, while another modified syringe pump acts as a one-dimensional translation stage, moving the needle across the membrane much like a pen. This method consumes very small volumes of reagents, and is a viable option for laboratory-scale fabrication and prototyping of point-of-care rapid diagnostic test strips.
- Published
- 2010
- Full Text
- View/download PDF
4. The extraction of liquid, protein molecules and yeast cells from paper through surface acoustic wave atomization.
- Author
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Qi A, Yeo L, Friend J, and Ho J
- Subjects
- Animals, Cattle, Cell Separation instrumentation, Electrophoresis, Microfluidic Analytical Techniques, Ovalbumin isolation & purification, Serum Albumin, Bovine isolation & purification, Surface Properties, Acoustics, Cell Separation methods, Paper, Proteins isolation & purification, Yeasts cytology
- Abstract
Paper has been proposed as an inexpensive and versatile carrier for microfluidics devices with abilities well beyond simple capillary action for pregnancy tests and the like. Unlike standard microfluidics devices, extracting a fluid from the paper is a challenge and a drawback to its broader use. Here, we extract fluid from narrow paper strips using surface acoustic wave (SAW) irradiation that subsequently atomizes the extracted fluid into a monodisperse aerosol for use in mass spectroscopy, medical diagnostics, and drug delivery applications. Two protein molecules, ovalbumin and bovine serum albumin (BSA), have been preserved in paper and then extracted using atomized mist through SAW excitation; protein electrophoresis shows there is less than 1% degradation of either protein molecule in this process. Finally, a solution of live yeast cells was infused into paper, which was subsequently dried for preservation then remoistened to extract the cells via SAW atomization, yielding live cells at the completion of the process. The successful preservation and extraction of fluids, proteins and yeast cells significantly expands the usefulness of paper in microfluidics.
- Published
- 2010
- Full Text
- View/download PDF
5. The extraction of liquid, protein molecules and yeast cells from paper through surface acoustic wave atomizationElectronic supplementary information (ESI) available: SAW atomisation for paper extraction. See DOI: 10.1039/b915833b.
- Author
-
Aisha Qi, Leslie Yeo, James Friend, and Jenny Ho
- Subjects
- *
EXTRACTION (Chemistry) , *PROTEINS , *YEAST , *PAPER , *MICROFLUIDIC devices , *ACOUSTIC surface wave devices , *ATOMIZATION , *MASS spectrometry - Abstract
Paper has been proposed as an inexpensive and versatile carrier for microfluidics devices with abilities well beyond simple capillary action for pregnancy tests and the like. Unlike standard microfluidics devices, extracting a fluid from the paper is a challenge and a drawback to its broader use. Here, we extract fluid from narrow paper strips using surface acoustic wave (SAW) irradiation that subsequently atomizes the extracted fluid into a monodisperse aerosol for use in mass spectroscopy, medical diagnostics, and drug delivery applications. Two protein molecules, ovalbumin and bovine serum albumin (BSA), have been preserved in paper and then extracted using atomized mist through SAW excitation; protein electrophoresis shows there is less than 1% degradation of either protein molecule in this process. Finally, a solution of live yeast cells was infused into paper, which was subsequently dried for preservation then remoistened to extract the cells viaSAW atomization, yielding live cells at the completion of the process. The successful preservation and extraction of fluids, proteins and yeast cells significantly expands the usefulness of paper in microfluidics. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
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