Protein Analysis At The Single Cell Level By Nonlinear Laser Wave Mixing Spectroscopy For High Throughput Capillary Electrophoresis Applications

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Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications

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ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (656 download)

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Book Synopsis Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications by :

Download or read book Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications written by and published by . This book was released on 2004 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: SADRI, BEHROKH BAGHERIFAR. Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications. (Under the direction of William G. Tong and Morteza G. Khaledi) Nonlinear degenerate four-wave mixing is presented as an ultrasensitive optical absorption-based method for detection and measurement of biological samples. Wave-mixing imaging detection technique can localize and quantify biomolecules in single cells and tissue sections with excellent spatial distribution of light absorbed by a target sample. Cellular components can be label-free or labeled with a chromophore or a fluorophore and imaged by wave mixing using a CCD camera. In a 2-D forward-scattering wave-mixing geometry, two overlapping laser beams form interference gratings and transfer their energy to an absorbing medium, creating thermal gratings followed by changes in the refractive index. The probe beam diffracts off these laser- induced gratings to produce the signal beam, which is detected by a CCD camera or a photodiode. A single bio cell can be placed in a glass slide and as the laser beams probe the labeled cellular component, the CCD camera captures wave-mixing signals corresponding to the absorbing cellular components. This nonlinear imaging technique can be used for both live and fixed cells in real time to obtain information on sequential changes in the number, morphology and distribution of cellular components in a single cell. Nonlinear laser wave-mixing spectroscopy coupled with capillary electrophoresis provides a novel ultrasensitive method for single-cell protein analysis. This method is used to detect proteins separated within a single cell. Nonlinear wave mixing has many advantages including quadratic dependency on analyte concentration, high spatial resolution and small sample requirements. Furthermore, wave mixing offers excellent detection sensitivity levels even when using very short optical path lengths, an.

Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications

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ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (656 download)

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Book Synopsis Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications by :

Download or read book Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications written by and published by . This book was released on 2010 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: SADRI, BEHROKH BAGHERIFAR. Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary Electrophoresis Applications. (Under the direction of William G. Tong and Morteza G. Khaledi) Nonlinear degenerate four-wave mixing is presented as an ultrasensitive optical absorption-based method for detection and measurement of biological samples. Wave-mixing imaging detection technique can localize and quantify biomolecules in single cells and tissue sections with excellent spatial distribution of light absorbed by a target sample. Cellular components can be label-free or labeled with a chromophore or a fluorophore and imaged by wave mixing using a CCD camera. In a 2-D forward-scattering wave-mixing geometry, two overlapping laser beams form interference gratings and transfer their energy to an absorbing medium, creating thermal gratings followed by changes in the refractive index. The probe beam diffracts off these laser- induced gratings to produce the signal beam, which is detected by a CCD camera or a photodiode. A single bio cell can be placed in a glass slide and as the laser beams probe the labeled cellular component, the CCD camera captures wave-mixing signals corresponding to the absorbing cellular components. This nonlinear imaging technique can be used for both live and fixed cells in real time to obtain information on sequential changes in the number, morphology and distribution of cellular components in a single cell. Nonlinear laser wave-mixing spectroscopy coupled with capillary electrophoresis provides a novel ultrasensitive method for single-cell protein analysis. This method is used to detect proteins separated within a single cell. Nonlinear wave mixing has many advantages including quadratic dependency on analyte concentration, high spatial resolution and small sample requirements. Furthermore, wave mixing offers excellent detection sensitivity levels even when using very short optical path lengths, an.

Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection

Author : Michael D. Harvey
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection by : Michael D. Harvey

Download or read book Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection written by Michael D. Harvey and published by . This book was released on 2001 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The trend towards high throughput applications and miniaturization necessitates approaches capable of microlitre volume sampling and low protein concentration detection. Furthermore, one of the major trends in high throughput screening is the growing replacement of technologies that depend on radioactivity to generate a signal with those that rely on fluorescence. This trend towards non-radioactive detection in general can be understood by some of the advantages inherent to these methods over radioactive modes. These include a significant reduction in safety concerns leading to a relaxation of strict laboratory procedures, elimination of expensive waste disposal, extended shelf-life of labeled reagents, and the possibility of acquiring multiplexed data through the spectral isolation of different wavelength signals. A variety of capillary electrophoretic (CE) approaches utilizing laser-induced fluorescence (LIF) have thus been developed, providing researchers with valuable tools in protein analysis. Various covalent and non-covalent fluorescent derivatization approaches have been investigated, with emphasis on biochemical and/or clinical applications. The non-covalent dye, NanoOrange, is used as a clinical diagnostic tool for early disease diagnosis, quantitating nanomolar concentrations of human serum albumin in solution, and obtaining fluorescence-based biofluid profiles. An alternate non-covalent labeling approach utilizing the fluorescent probe, Sypro Red, and capillary gel electrophoresis allows for rapid, sensitive analysis of protein sample purity as well as molecular weight determination. These two non-covalent approaches are complemented by the development of a fluorescent Insulin-Like Growth Factor-I (IGF-I) analog for use in bioanalytical applications. Specific derivatization reaction conditions were developed to selectively label the N-terminus of the analog hence preserve biological activity. High-performance liquid chromatography and electrospray mass spectrometry were used to confirm the extent of labeling and modification site. Antibody recognition of this fluorescent analog was evaluated using CE-LIF, illustrating the clinical utility of this diagnostic reagent. In addition to the above CE-LIF approaches, a fourth capillary electrophoretic tool is provided for the clinical chemist. Rapid analysis of biofluids is of significant importance in early disease diagnosis. As such, an extensive CE-based analysis of human seminal plasma is presented. Separation conditions, sample stability, and protein/non-protein zone identification issues are addressed. This study and the CE-LIF methodologies discussed above represent original approaches to nanoscale protein analysis.

Advancing Capillary Electrophoresis-mass Spectrometry for Top-down Proteomics

Author : Tian Xu
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.3/5 (776 download)

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Book Synopsis Advancing Capillary Electrophoresis-mass Spectrometry for Top-down Proteomics by : Tian Xu

Download or read book Advancing Capillary Electrophoresis-mass Spectrometry for Top-down Proteomics written by Tian Xu and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Top-down proteomics (TDP) enables the proteome profiling of biological subjects at the proteoform level and understanding of differential functions associated with proteoform heterogeneity, such as sequence variation, post-translational modifications (PTMs), etc. Drastic advances on TDP technologies (e.g. sample preparation, separation/fractionation, fragmentation, bioinformatics, etc.) have been achieved in the past decades. Further improvements in separation remain desired for better analysis throughput and deeper proteome coverage. Capillary electrophoresis (CE), including capillary zone electrophoresis (CZE) and capillary isoelectric focusing (cIEF), provide superior separation performance for proteoforms. This dissertation focuses on the advancement of CE-MS-based tools on throughput, separation resolution, and capacity for TDP and utility of these tools for biological applications.In Chapter 2, we developed high-throughput and high-capacity cIEF-MS/MS platforms. The high-throughput platform enables efficient identification and quantification of proteoforms (less than one hour per run), whereas the high-capacity cIEF-MS/MS provides large number of proteoform identifications (IDs, more than 700 proteoforms in a single shot analysis) which is valuable for deep TDP. In Chapter 3, we further improved the stability and robustness of cIEF-MS platform using optimized linear polyacrylamide (LPA) capillary coating and catholyte with lower pH (pH~10). The work achieved high-resolution characterization and accurate isoelectric point (pI) determination of charge variants (~0.1 pI difference) of monoclonal antibodies (mAbs). In Chapter 4, we developed a nondenaturing cIEF-MS platform for ultrahigh resolution characterization of microheterogeneity of a variety of protein complexes. Typically, pI determinations of variants in protein complexes allow us to decipher how sequence or PTM variations modulate the pIs of the protein complexes. In Chapter 5, while CZE-MS/MS is a well-developed approach, for the first time, we coupled FAIMS to CZE-MS/MS to facilitate online gas-phase fractionation of proteoforms. The FAIMS greatly enhanced the sensitivity of the system and expanded the number of proteoform IDs, especially large proteoform IDs. The work renders CZE-FAIMS-MS/MS as a new powerful multidimensional platform for deep TDP.In Chapters 6 and 7, we applied cIEF-MS/MS and CZE-MS/MS for studying the sexual dimorphism of zebrafish brains and proteoform-level differences between metastatic and nonmetastatic colorectal cancer (CRC) cells, respectively. In Chapter 6, quantitative TDP of thousands of proteoforms from male and female zebrafish brains by cIEF-MS/MS based approach discovered various overexpressed proteoforms in male or female brains that are closely associated with hormone activity. In Chapter 7, We performed deep TDP study of non-metastatic and metastatic CRC cells (SW480 and SW620) using CZE-MS/MS based multidimensional platform and identified more than 20,000 proteoforms of over 2,000 proteins from the two cell lines, which presents around 5-folds higher number of proteoform IDs in comparison with previous TDP studies of human cancer cells. The work revealed significant discrepancies between the two isogenic cell lines regarding proteoform and single amino acid variant (SAAV) profiles. Quantitative data disclosed differentially expressed proteoforms between the two cell lines and their corresponding genes were connected to cancer pathways and networks.

Degenerate Four-Wave Mixing Interfaced with Capillary Electrophoresis as a Bioanalytical Method for Small Molecules, Peptides, and Proteins

Download Degenerate Four-Wave Mixing Interfaced with Capillary Electrophoresis as a Bioanalytical Method for Small Molecules, Peptides, and Proteins PDF Online Free

Author : Manna Frances Iwabuchi
Publisher :
ISBN 13 :
Total Pages : 187 pages
Book Rating : 4.:/5 (911 download)

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Book Synopsis Degenerate Four-Wave Mixing Interfaced with Capillary Electrophoresis as a Bioanalytical Method for Small Molecules, Peptides, and Proteins by : Manna Frances Iwabuchi

Download or read book Degenerate Four-Wave Mixing Interfaced with Capillary Electrophoresis as a Bioanalytical Method for Small Molecules, Peptides, and Proteins written by Manna Frances Iwabuchi and published by . This book was released on 2015 with total page 187 pages. Available in PDF, EPUB and Kindle. Book excerpt: Degenerate four-wave mixing (DFWM) is demonstrated as an ultrasensitive detection method for the study of neurodegenerative disease-related biomolecules in a capillary electrophoresis system. Nonlinear absorption-based wave mixing offers important advantages including high spatial resolution, small probe volumes down to picoliter, small sample requirements, effective signal collections, and low background noise levels. The laser wave mixing is easily interfaced with microcapillaries for a continuous-flow mode detection and a capillary electrophoresis-mode detection. The optimal conditions for laser wave mixing and capillary electrophoresis is investigated to perform ultrasensitive detection and molecular weight-based separation for proteins. A cost-effective fluorophore is employed to target analytes, bovine serum albumin (BSA), antibody of HIV-1 capsid protein p24, and monoclonal antibody of breast cancer marker CA15-3. Concentration (and mass) detection limits of 1.4 x 10−10 M (1.0 x 10−20 mol), 6.6 x 1010−10 M (5.1 x 10−20 mol), and 6.7 x 10−12 M (5.2 x 10−22 mol) are determined for BSA, p24 antibody, and CA15-3 antibody. For the first time, [alpha]-synuclein is analyzed using fluorescein isothiocyanate (FITC), QSY35 acetic acid succinimidyl ester, and ChromeoTM P503 by wave mixing-based capillary zone electrophoresis and capillary sieving electrophoresis. Detection limits of 1.4 x 10−13 M (1.1 x 10−23 mol), 1.4 x 10−10 M (1.1 x 1010−20 mol), and 3.8 x 10−13 M (3.0 x 10−23 mol) are determined for FITC, QSY35, and ChromeoTM P503-conjugated [alpha]-synuclein. Tetramer [alpha]-synuclein is observed to be the most abundant species in the buffer system in which separation is executed. Native label-free adenosine and dopamine, and chromophore-conjugated glutamate and [gamma]-aminobutyric acid are detected at ultrasensitive concentration levels using wave mixing-based micellar electrokinetic chromatography. Detection limits of 1.0x10−13 M (5.6x10−24 mol corresponds to 4 molecules), 1.2x10−9 M (6.6x10−20 mol), 3.7x10−10 M (2.9x10−20 mol), and 4.1x10−11 M (3.2x10−21 mol) are determined for native label-free adenosine and dopamine, dabsylated glutamate and [gamma]-aminobutyric acid. [Beta]-Amyloid 1-42 and 1-40 peptides are investigated using wave mixing based capillary zone electrophoresis. Detection limits of 8.2 x 10−13 M (6.5 x 10−23 mol) and 1.2 x 10−12 M (9.5 x 10−23 mol) are determined for A[Beta]1-42 and A[Beta]1-40.

Capillary Electrophoresis of Proteins and Peptides

Author : Nguyet Thuy Tran
Publisher : Humana
ISBN 13 : 9781493981533
Total Pages : 234 pages
Book Rating : 4.9/5 (815 download)

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Book Synopsis Capillary Electrophoresis of Proteins and Peptides by : Nguyet Thuy Tran

Download or read book Capillary Electrophoresis of Proteins and Peptides written by Nguyet Thuy Tran and published by Humana. This book was released on 2018-05-30 with total page 234 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book provides a comprehensive survey of recent developments and applications of high performance capillary electrophoresis in the field of protein and peptide analysis with a distinct focus on the analysis of intact proteins. With practical detail, the contents cover different modes of capillary electrophoresis (CE) useful for protein and peptide analysis, CZE, CIEF, ACE, CGE, and different types of application such as the quality control of therapeutic proteins and monoclonal antibodies, clinical analyses of chemokines in tissues, qualitative and quantitative analysis of vaccine proteins, and determination of binding constants in complexes involving peptides or proteins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and exhaustive, Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols serves both beginners and experts with a collection of the current and most active topics in this vital field of study.

Single Cell Protein Analysis

Author : Anup K. Singh
Publisher :
ISBN 13 : 9781493929870
Total Pages : 254 pages
Book Rating : 4.9/5 (298 download)

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Book Synopsis Single Cell Protein Analysis by : Anup K. Singh

Download or read book Single Cell Protein Analysis written by Anup K. Singh and published by . This book was released on 2015 with total page 254 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume highlights recent developments in flow cytometry, affinity assays, imaging, mass spectrometry, microfluidics and other technologies that enable analysis of proteins at the single cell level. The book also includes chapters covering a suite of biochemical and biophysical methods capable of making an entire gamut of proteomic measurements, including analysis of protein abundance or expression, protein interaction networks, post-translational modifications, translocation and enzymatic activity.

Capillary Electrophoresis-Mass Spectrometry

Author : James Q. Xia
Publisher : Springer
ISBN 13 : 9783319834757
Total Pages : 74 pages
Book Rating : 4.8/5 (347 download)

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Book Synopsis Capillary Electrophoresis-Mass Spectrometry by : James Q. Xia

Download or read book Capillary Electrophoresis-Mass Spectrometry written by James Q. Xia and published by Springer. This book was released on 2018-06-28 with total page 74 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the latest developments in capillary electrophoresis-mass spectrometry for the analysis of therapeutic proteins. The application of capillary electrophoresis-mass spectrometry (CE-MS) coupling technology in the analysis of recombinant therapeutic proteins is detailed thoroughly. Specific topics include recent developments in coupling capillary electrophoresis with mass spectrometry for the quality control of monoclonal antibody therapeutics, top-down analysis of monoclonal antibody using the CE-MS platform, and detection of host cell protein impurities. Comprehensive characterization of antibody-drug conjugates (ADCs) by coupling capillary electrophoresis with mass spectrometry is also covered. This is an ideal book for scientists in the life science and biopharmaceutical industry who are working on characterizing the PTMs of monoclonal antibodies, as well as graduate students and researchers in the separation science and biological mass spectrometry fields.

Microcharacterization of Proteins

Author : Roland Kellner
Publisher : John Wiley & Sons
ISBN 13 : 3527613978
Total Pages : 346 pages
Book Rating : 4.5/5 (276 download)

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Book Synopsis Microcharacterization of Proteins by : Roland Kellner

Download or read book Microcharacterization of Proteins written by Roland Kellner and published by John Wiley & Sons. This book was released on 2008-09-26 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteomics - the analysis of the whole set of proteins and their functions in a cell - is based on the revolutionary developments which have been achieved in protein analysis during the last years. The number of finished genome projects is growing and in parallel there is a dramatically increasing need to identify the products of revealed genes. Acting on a micro level modern protein chemistry increases our understanding of biological events by elucidating the relevant structure-function relationships. The second edition of the successful title Microcharacterization of Proteins presents a current overview of modern protein analysis: From sample preparation to sequence analysis, mass spectrometry and bioinformatics it informs about the tools needed in protein research. This makes the book indispensable for everyone involved in proteomics!

Capillary Electrophoresis-mass Spectrometry for Top-down Protein Analysis

Author : Alexander Stolz
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (137 download)

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Book Synopsis Capillary Electrophoresis-mass Spectrometry for Top-down Protein Analysis by : Alexander Stolz

Download or read book Capillary Electrophoresis-mass Spectrometry for Top-down Protein Analysis written by Alexander Stolz and published by . This book was released on 2023* with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Due to the high sensitivity, selectivity, and the possibility for detailed molecular characterisation, mass spectrometry (MS) is the analytical method of choice for top-down protein biomarker discovery. Due to the low sample consumption, high separation efficiency, and the unique and complementary selectivity, capillary zone electrophoresis (CZE)-MS represents an interesting alternative to the traditionally used liquid chromatography (LC)-MS platforms. In this thesis, instrumental and methodological concepts were developed to increase the potential of CZE-MS for intact proteins. The first part of the thesis describes the development of a nanoflow sheath liquid interface for the efficient coupling of CZE and MS. The interface was developed with a focus on fast setup, easy handling and analytical robustness and has been used for most applications in this thesis. Furthermore, a CZE-MS screening platform for the identification and characterisation of known and unknown Hb variants from DBS samples was developed. The application of SMIL coatings enables efficient separation of closely-related proteoforms and even positional isomers of glycated Hb on the intact level. In the last part of the thesis, nanoLC and CZE-MS were coupled in a heart-cut approach using a polymer nanoliter valve. The platform was used for the glycosylation profiling of heterogeneous alpha-1 acid glycoprotein (AGP). This approach enables the assignment of notably more glycoforms from a lower concentrated AGP sample, compared to CZE-MS alone. In a proof-of-concept study, the platform was further extended to operate in the selective comprehensive mode. With a single injection, 19% more glycoforms were assigned compared to the heart-cut approach with 3 injections. The here presented instrumental and methodological concepts show the great potential of CZE-MS in the context of clinical protein analysis. Especially the combination of LC and CZE in multidimensional separation platforms shows great potential.

High-throughput Analysis and Protein Engineering Using Microcapillary Arrays

Author : Bob Chen
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (936 download)

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Book Synopsis High-throughput Analysis and Protein Engineering Using Microcapillary Arrays by : Bob Chen

Download or read book High-throughput Analysis and Protein Engineering Using Microcapillary Arrays written by Bob Chen and published by . This book was released on 2015 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Over the past decade, high-throughput technologies have allowed researchers to gain unprecedented insights into intrinsically complex and interconnected biological systems. As examples, whole-genome sequencing has enabled the identification of crucial genes and mutations underlying disease pathophysiology, DNA microarrays have been used to elucidate transcription patterns involved in healthy and diseased states, and large-scale proteomics methods have helped map the connectivity of cell signaling networks that orchestrate responses to growth factors and other external stimuli. In contrast, analogously powerful approaches for rapidly and deeply interrogating the sequence-structure-activity relationship of proteins, with functional read-outs that span a range of biophysical and biochemical measurements, have lagged because of technical challenges. In this dissertation, I describe the development of a new technology platform, termed μSCALE (microcapillary single-cell analysis and laser extraction), that aims to improve high-throughput protein characterization and engineering. The μSCALE platform is built around a microcapillary array, automated fluorescence imaging, and a novel laser-based sample recovery method. The platform spatially segregates millions of cells or their protein products in a microcapillary array and interrogates the dense array within minutes. μSCALE also enables recovery of target sample from the microcapillary array using a precise laser-based extraction technique. To showcase capabilities and breadth of μSCALE, I applied it to three distinct protein-analysis and engineering applications, using libraries expressed in yeast or bacteria. In each case, I demonstrated feasibility and then engineered a novel protein variant, identifying new binders against a clinical target, creating a novel fluorescent protein biosensor, or reducing inhibition of an enzyme. Compared to conventional screening techniques, such as FACS and plate-based screens, the μSCALE affords several distinctions advantages, including a five orders of magnitude improvement in throughput, direct cell imaging, and high sorting purity and yield. Collectively, these benefits present μSCALE as an exciting, novel technology to the field of protein engineering and beyond.

Protein Analysis and Purification

Author : I.M. Rosenberg
Publisher : Springer Science & Business Media
ISBN 13 : 1475711085
Total Pages : 453 pages
Book Rating : 4.4/5 (757 download)

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Book Synopsis Protein Analysis and Purification by : I.M. Rosenberg

Download or read book Protein Analysis and Purification written by I.M. Rosenberg and published by Springer Science & Business Media. This book was released on 2013-03-14 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

Protein Analysis using Mass Spectrometry

Author : Mike S. Lee
Publisher : John Wiley & Sons
ISBN 13 : 1118605195
Total Pages : 290 pages
Book Rating : 4.1/5 (186 download)

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Book Synopsis Protein Analysis using Mass Spectrometry by : Mike S. Lee

Download or read book Protein Analysis using Mass Spectrometry written by Mike S. Lee and published by John Wiley & Sons. This book was released on 2017-06-19 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents Practical Applications of Mass Spectrometry for Protein Analysis and Covers Their Impact on Accelerating Drug Discovery and Development Covers both qualitative and quantitative aspects of Mass Spectrometry protein analysis in drug discovery Principles, Instrumentation, Technologies topics include MS of peptides, proteins, and ADCs , instrumentation in protein analysis, nanospray technology in MS protein analysis, and automation in MS protein analysis Details emerging areas from drug monitoring to patient care such as Identification and validation of biomarkers for cancer, targeted MS approaches for biomarker validation, biomarker discovery, and regulatory perspectives Brings together the most current advances in the mass spectrometry technology and related method in protein analysis

Capillary Electrophoresis with Multiple Readout Techniques for Protein Analysis

Author :
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (93 download)

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Book Synopsis Capillary Electrophoresis with Multiple Readout Techniques for Protein Analysis by :

Download or read book Capillary Electrophoresis with Multiple Readout Techniques for Protein Analysis written by and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Protein and Peptide Analysis by LC-MS

Author : Thomas Letzel
Publisher : Royal Society of Chemistry
ISBN 13 : 1849733147
Total Pages : 195 pages
Book Rating : 4.8/5 (497 download)

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Book Synopsis Protein and Peptide Analysis by LC-MS by : Thomas Letzel

Download or read book Protein and Peptide Analysis by LC-MS written by Thomas Letzel and published by Royal Society of Chemistry. This book was released on 2011-07-22 with total page 195 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is the first example in presenting LC-MS strategies for the analysis of peptides and proteins with detailed information and hints about the needs and problems described from experts on-the-job. The best advantage is -for sure- the practical insight of experienced analysts into their novel protein analysis techniques. Readers starting in 'Proteomics' should be able to repeat each experiment with own equipment and own protein samples, like clean-up, direct protein analysis, after (online) digest, with modifications and others. Furthermore, the reader will learn more about strategies in protein analysis, like quantitative analysis, industrial standards, functional analysis and more.

Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry

Download Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry PDF Online Free

Author : Feng Zhou
Publisher : ProQuest
ISBN 13 : 9780549388937
Total Pages : pages
Book Rating : 4.3/5 (889 download)

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Book Synopsis Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry by : Feng Zhou

Download or read book Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry written by Feng Zhou and published by ProQuest. This book was released on 2008 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: With the development of genomics, separation technology and mass spectrometry, proteomics has become an important tool in biological research. Characterization of posttranslational modification (PTM) by mass spectrometry is attracting an increasing attention because of its importance in signal transduction and the difficulty in obtaining this information from genomic methods. Bottom-up methods, which analyze the peptides from enzymatic digestion of proteins, are widely used for proteomic research. However, bottom-up methods may lose some important information when characterizing PTM since not all peptides are detected. In contrast, top-down methods directly characterize intact proteins, which makes it possible to retain PTM information. This dissertation focuses on the characterization of proteins by coupling high throughput methods, capillary isoelectric focusing electrophoresis (CIEF) and electrospray ionization (ESI) mass spectrometry (MS). In this work, a CIEF-RPLC-ESIMS system is designed and constructed for 2-D profiling of protein mixture. The microdialysis membrane based cathodic cell used in this system not only serves as the cathodic cell in CIEF separation, but also transfers the focused proteins separated by CIEF to RPLC. After eliminating the interference of ampholyte, a few fmol of protein can be detected with this system. The detection limit of this system is one order of magnitude lower than 2-D PAGE. By taking the advantage of high resolving power with mass spectrometry and multidimensional separation, the theoretical peak capacity of this system can reach 1x10 6, which is two orders of magnitude higher than theoretical peak capacity of 2-D PAGE. This system is firstly tested with a standard protein mixture. After slight modification, this system is used to profile the intact protein from a yeast enzyme concentrate and a lysate of the green sulfur bacterium Chlorobium tepidum. The pI vs. MW profile obtained from CIEF-RPLC-MS compares favorably with theoretical data derived from C. tepidum genome and experimental data obtained from 2-D-PAGE.

Protein Sequencing and Identification Using Tandem Mass Spectrometry

Author : Michael Kinter
Publisher : John Wiley & Sons
ISBN 13 : 0471231886
Total Pages : 321 pages
Book Rating : 4.4/5 (712 download)

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Book Synopsis Protein Sequencing and Identification Using Tandem Mass Spectrometry by : Michael Kinter

Download or read book Protein Sequencing and Identification Using Tandem Mass Spectrometry written by Michael Kinter and published by John Wiley & Sons. This book was released on 2005-04-12 with total page 321 pages. Available in PDF, EPUB and Kindle. Book excerpt: How to design, execute, and interpret experiments for protein sequencing using mass spectrometry The rapid expansion of searchable protein and DNA databases in recent years has triggered an explosive growth in the application of mass spectrometry to protein sequencing. This timely and authoritative book provides professionals and scientists in biotechnology research with complete coverage of procedures for analyzing protein sequences by mass spectrometry, including step-by-step guidelines for sample preparation, analysis, and data interpretation. Michael Kinter and Nicholas Sherman present their own high-quality, laboratory-tested protocols for the analysis of a wide variety of samples, demonstrating how to carry out specific experiments and obtain fast, reliable results with a 99% success rate. Readers will get sufficient experimental detail to apply in their own laboratories, learn about the proper selection and operation of instruments, and gain essential insight into the fundamental principles of mass spectrometry and protein sequencing. Coverage includes: * Peptide fragmentation and interpretation of product ion spectra * Basic polyacrylamide gel electrophoresis * Preparation of protein digests for sequencing experiments * Mass spectrometric analysis using capillary liquid chromatography * Techniques for protein identification by database searches * Characterization of modified peptides using tandem mass spectrometry And much more