New Sample Fractionation Strategies for Proteomic Analyses by LC MS
|
|
- Peregrine Lewis
- 6 years ago
- Views:
Transcription
1 16 Current Trends in Mass Spectrometry November 2006 New Sample Fractionation Strategies for Proteomic Analyses by LC MS Mass spectrometry has long been a preferred tool for protein identification and biomarker discovery, but preparation of biological samples remains a challenge. Hindrances include the wide range of protein concentrations, sample complexity, and loss or alteration of important proteins due to sample handling. This article describes recent developments in sample fractionation technologies that are overcoming these challenges in interesting ways and are enabling in-depth proteomic studies that were not possible in the past. Jerome Bailey, Peter Mrozinski, Tobias Preckel, Christine Miller, James Martosella, and Robert Kincaid Proteomic analyses based upon liquid chromatography mass spectrometry (LC MS) are quite powerful, but when protein digests are too complex, lowerconcentration peptides often escape detection. This is where sample preparation plays a key role. For almost any proteomic analysis today, robust and reproducible fractionation techniques are the key to ensuring that MS-MS can identify the maximum number of proteins. At the same time, to avoid inadvertent loss or alteration of critical proteins, it is important that fractionation steps minimize the number of sample manipulations. Successful fractionation of biological samples is crucial for identification of diagnostic markers and therapeutic targets. Some biological samples are excellent candidates for biomarker study, but researchers have not yet fully explored them because of difficulties with sample preparation. Examples include plasma and serum analyses, in which highly abundant proteins can mask the low-abundance proteins of interest, and membrane-protein analysis, in which hydrophobicity contributes to losses during sample handling. With new developments in sample fractionation techniques, these previously inaccessible samples are becoming accessible. The Challenge of Plasma Because plasma comes into contact with almost all of the tissues in the body and is easily obtainable, it is an important source of new protein biomarkers for diagnostics and drug discovery and development. However, plasma is a very complex sample that presents unique challenges, including the fact that proteins are present over a wide concentration range from micrograms or milligrams per milliliter for the most abundant proteins down to nanograms or picograms per milliliter for the less abundant species. Seven of the most abundant proteins in plasma albumin, IgG, transferrin, haptoglobin, IgA, anti-trypsin, and fibrinogen make up about 90% of the total protein mass. These proteins must be removed to enable detection of low-concentration biomarkers by LC MS. Another challenge with plasma is the complexity of the sample that remains after removal of the most abundant proteins. Sample complexity is an obstacle that is common to many proteomic samples. The interesting proteins must be fractionated to produce simpler mixtures with fewer coeluted species. Only then is it possible to maximize the number of proteins that can be identified in the search for important protein biomarkers. Reproducible Removal of High-Abundance Proteins Multiple affinity removal (immunodepletion) is an excellent method for eliminating highly abundant proteins. With this technique, the sample is passed through a column or other medium, where antibody antigen interactions remove tar-
2 18 Current Trends in Mass Spectrometry November 2006 Normalized absorbance (mau) Run 1 Run 40 Run 80 Run 120 Run 160 Run 200 Flow-through fraction Bound fraction Time (min) Figure 1: Overlay of chromatograms from automated runs 1, 40, 80, 120, 160, and 200 from an Agilent Human Plasma 7 Multiple Affinity Removal column. geted proteins. Immunodepletion requires optimized buffers that minimize binding of nontargeted proteins, and that ensure reproducibility and long column life. The method requires one buffer to load, wash, and regenerate the column, and a second buffer to elute the bound high-abundance proteins. Several factors contribute to successful immunodepletion. First and most obvious, the process must be very selective. In other words, it must remove a very high percentage of the undesired proteins while allowing the other proteins to pass through. The removal of abundant proteins can be optimized by using antibodies that are specific to both the protein and the species of interest, and by mixing them in the proper (biological) ratios. Well-designed columns can remove at least 98% of seven targeted high-abundance proteins, as demonstrated by enzyme-linked immunosorbent assays (ELISA). To maintain high recoveries of biomarkers, it is equally important to minimize the nonspecific binding of interesting low-abundance proteins to the antibodies. The immunodepletion process also must circumvent a tricky problem the association of many biomarkers with sticky carrier proteins such as albumin. To avoid removing these biomarkers along with the albumin, it is important that buffers minimize protein protein interactions. To enable detection of low-abundance proteins by LC MS, it is very important for the affinity removal column to have sufficient sample capacity. Currently, commercially available 100 mm 4.6 mm columns for human fluids allow loading of 70 µl of plasma, while 100 mm 10 mm columns allow loading of 300 µl of plasma. This high loadability permits LC MS-MS detection of biomarkers in the nanogram-permilliliter range. Finally, the multiple affinity removal process should be easily automated, fast, and reproducible. Figure 1 shows an overlay of six chromatograms obtained during the automated removal of seven highly abundant proteins from 200 human plasma samples. A conventional LC system was used to automate the sample injection, as well as the introduction of the two buffers that the immunodepletion process requires. The figure shows reproducible results for over 200 injections. In addition, one-dimensional gel patterns of the flow-through fractions from multiple runs (data not shown) indicated consistency and robustness of the method. New Methods for Protein-Level Fractionation To achieve the greatest number of protein identifications with LC MS analyses, samples typically must be fractionated in multiple dimensions. Fractionation can occur either before or after protein digestion. At the peptide level, researchers often successfully reduce sample complexity via a combination of strong cation exchange and reversed-phase LC. Fractionation can occur instead (or in addition) at the protein level. Macroporous reversed-phase C18 (mrp- C18, Agilent Technologies) columns constitute one relatively new way to achieve reproducible, high-resolution separations of proteins with high recoveries. OFFGEL electrophoresis (pi-based fractionation system developed jointly by Agilent and DiagnoSwiss S.A., Monthey, Switzerland) is another new technique for protein fractionation (1,2). It separates either proteins or peptides by isoelectric point (pi), with sample recovery in the solution phase. The solution-phase, pibased fractionation system has the advantage of providing the protein or peptide pi, which scientists can then use to validate the results of protein database searches that are based upon MS-MS data. Both macroporous reversed-phase C18 columns and the solution-phase, pi-based
3 November 2006 Current Trends in Mass Spectrometry Control serum Absorbance (mau) Cortisol-deficient serum Serum with elevated rheumatoid factor Time (min) Figure 2: Chromatograms from separation of immunodepleted serum on an Agilent mrp-c18 column. fractionation system allow one to load sufficient sample so that low-level proteins can be detected in subsequent LC MS analyses. These techniques are discussed in detail below. Columns for High Protein Recoveries The use of macroporous reversed-phase C18 columns is a relatively new method for protein fractionation. These columns exhibit excellent resolution, loadability, and reproducibility. Conventional reversed-phase LC columns often lack the resolution needed for complex samples like cell lysates, and protein recoveries are typically only 30 80%. The macroporous columns provide much better resolution. They also dramatically improve recoveries because a proprietary surface treatment, in combination with the large-pore bonded-silica material, prevents irreversible protein adsorption. Under optimized conditions, protein recoveries are typically greater than 95% and have been measured at 98% for immunodepleted serum (3). High recoveries eliminate sample cross-contamination and allow direct sample comparison, as shown in Figure 2. In addition, macroporous reversedphase columns are ideal for desalting and concentrating protein samples (4). The process is faster than with spin concentrators and avoids the molecular weight cutoff that typically is associated with that technique. Immunodepletion in Combination with Macroporous Reversed- Phase C18 Fractionation For biomarker discovery in human plasma, it is very powerful to combine multiple affinity removal of highly abundant proteins (discussed earlier) with macroporous reversed-phase fractionation of the remaining proteins. The transition between the two techniques is very easy; after denaturation with urea, researchers apply the immunodepleted plasma directly onto the macroporous reversed-phase C18 column (5). A recent study took advantage of the combination of immunodepletion and macroporous reversed-phase C18 fractionation (6). After affinity removal, the researchers concentrated, desalted, and fractionated the plasma with the macroporous reversed-phase C18 column at 80 C. The optimized LC conditions provided high protein recoveries, enhanced peak resolution, and reproducible fractionation. After macroporous reversed-phase fractionation, the researchers digested the proteins in solution with trypsin. They analyzed the peptides by LC MS-MS using a polymeric-based microfluidic device in combination with an ion trap. The overall workflow minimized sample manipulations; for example, it was not necessary to perform dialysis, buffer exchange, or precipitation. The combination of affinity removal with high-capacity macroporous reversed-phase C18 fractionation and subsequent peptide analysis allowed the identification of hundreds of low-abundance plasma proteins. Fractionation of Difficult Membrane Proteins The macroporous reversed-phase C18 columns have proven extremely useful for studies of membrane proteins. Although membrane proteins are an excellent source of potential biomarkers, they have received less study because their hydrophobicity poses recovery challenges. On both gels and most reversed-phase LC columns, they resolve poorly and are difficult to recover. Standard C18 reversed-phase columns irreversibly bind hydrophobic proteins, causing shifts in retention time and irreproducible peak areas. The latest well-engineered macroporous reversed-phase columns overcome these problems, making the macroporous reversed-phase C18 columns excellent tools for biomarker discovery with these difficult samples (7).
4 20 Current Trends in Mass Spectrometry November 2006 IPG gel IPG gel IPG gel Starts with micro-wells filled with sample diluted in buffer Protein A Protein A ph gradient ph gradient ph gradient A recent study showed the advantage of a macroporous reversed-phase column for fractionation of membrane proteins from HeLa cells (8). The workflow incorhv hv Protein B hv Protein B Electrode Electrode Electrode Figure 3: Schematic of solution-phase, pi-based protein and peptide fractionation. porated high-resolution fractionation steps but required a minimum of sample manipulations. The membrane proteins were solubilized and then were fractionated using an optimized gradient on a 50 mm 4.6 mm macroporous reversed-phase C18 column. After combining, 17 fractions underwent in-solution digestion. The peptides in the digests were fractionated with 11 salt steps on a capillary strong cation exchange column, and the resulting fractions were analyzed using the Agilent HPLC-Chip/MS system in combination with an ion trap. The MS-MS spectra were searched against the Swiss- Prot database (Swiss Institute of Bioinformatics, Geneva, Switzerland) using the Spectrum Mill MS Proteomics Workbench software (Agilent Technologies), and results were validated using conservative criteria. The result was identification of an unprecedented number of proteins. A total of 954 proteins were identified, including 470 membrane proteins, of which 337 were integral membrane proteins. A number of factors enabled the identification of such a large number of membrane proteins in this difficult sample. The unique surface treatment of the macroporous reversed-phase C18 column allowed very high protein recoveries. The column, in combination with the optimized gradient and elevated temperature, enabled excellent separation. Third, the high loadability of the macroporous reversed-phase C18 column, combined with the sensitivity of the peptide analysis system, allowed detection of numerous lowlevel peptides. pi-based Fractionation for Sample Recovery in the Liquid Phase The solution-phase, pi-based fractionation system is a relatively new separation sytem that fractionates either proteins or peptides by pi. It achieves the same resolution as immobilized ph gradient isoelectric focusing (IPG IEF), but it removes the requirement for tedious post-ief sample handling. After IEF, the IPG gel strips must be cut into sections and the peptides extracted and purified (9,10). With solution-phase, pi-based fractionation, the proteins or peptides end up in the liquid phase, which makes this technique directly compatible with LC MS. There is no need for additional sample extraction or purification steps that could lead to sample losses. Figure 3 shows the principle of solution-phase, pi-based fractionation. The
5 22 Current Trends in Mass Spectrometry November pl 10 Reversed-phase retention time (min) Figure 4: Visualization of color-coded protein concentrations from a HeLa cell lysate fractionated by a combination of pi (Agilent 3100 OFFGEL Fractionator, 24 fractions, ph 3 10) and reversed-phase LC (Agilent mrp-c18 column, 1536 fractions). sample is diluted in focusing buffer and is loaded into a series of isolated wells. The well forms a liquid-tight seal against a rehydrated conventional IPG gel strip. Upon application of an electric field, the proteins or peptides are forced to migrate along the electric field lines through the liquid-ipg gel strip system until they reach the well where the ph of the gel is equal to the pi of the peptide or protein. As the electric field extends into the wells, the majority of proteins or peptides are conveniently recovered in the liquid phase, which contains no detergents, surfactants, or other species that interfere with LC MS. One very useful aspect of solutionphase, pi-based fractionation is that it can be applied twice in the same experiment it can fractionate at both the protein level and the peptide level. Another convenient feature is that it requires a minimum of sample handling. Fractionated proteins can proceed directly to the digestion step, and fractionated peptides can proceed directly to LC MS. A unique advantage of solution-phase, pi-based fractionation is that researchers can adjust the separation by using IPG gels with various pi ranges. By using IPG gels with sufficient resolution (less than 0.2 pi units), it is possible to resolve isoforms of a protein (such as charged posttranslational modifications) and to recover them in individual solutions. Likewise, it is possible at the peptide level to identify amino acid mutations (11). Comparison of Solution-Phase, pi-based Fractionation with Strong Cation Exchange A recent experiment that compared fractionation of peptides with either strong cation exchange or solution-phase, pibased fractionation showed the advantage of solution-phase, pi-based fractionation for increased protein identifications. Immunodepleted human serum was digested tryptically. The resulting peptides were fractionated either into 24 fractions by solution-phase, pi-based fractionation (ph 3 to 10), or into 50 fractions by strong cation exchange. The individual fractions from both experiments were then analyzed using the Agilent HPLC-Chip/MS combined with an ion trap. The separation by solution-phase, pi-based fractionation produced twice the number of protein identifications and about three times the number of peptide identifications as the strong cation exchange technique. Use of pi Values from Solution- Phase, pi-based Fractionation for Validation of Database Search Results Another advantage of solution-phase, pibased fractionation is that the experimentally determined protein or peptide pis can be used as an additional validation tool when reviewing results of protein database searches. When the software used for a protein database search also calculates and displays peptide pis, it is easy to compare the calculated pis with those from the solution-phase, pi-based separation. Researchers can use the experimentally derived pis to rule out false positive matches, or they can validate additional matches with slightly lower scores when the pi values correspond as well. In a recent study, use of pi as an additional validation criterion led to 19 22% more peptide identifications (12). Very High-Resolution Fractionation for Complex Samples For complex samples such as cell lysates, it is possible to separate proteins first by
6 November 2006 Current Trends in Mass Spectrometry 23 solution-phase, pi-based fractionation and second by macroporous reversed-phase LC. The transition between the two techniques is seamless, with no sample manipulation in between. The combination can provide up to 1500 individual fractions, for an extremely high-resolution separation. With a macroporous reversed-phase LC column of smaller internal diameter, it is possible to introduce the fractions directly into a time-of-flight MS for measurement of molecular mass. Alternatively, with visualization software, such as the experimental software shown in Figure 4, it is possible to compare samples and pick selected fractions for digestion and MS- MS analysis. Relative to two-dimensional gel electrophoresis, the combination of solution-phase, pi-based fractionation and macroporous reversed-phase LC provides a more information-rich, faster, more reproducible, and easier-to-use alternative. Conclusion Successful execution of LC MS-based proteomic studies is highly dependent upon well-designed, seamless sample preparation schemes. For plasma and serum samples, affinity chromatography is proving to be a robust, reproducible method for removing highly abundant proteins that mask low-level proteins of interest. For many complex proteomic samples, fractionation at the protein level helps to reduce sample complexity so that it is possible to identify more proteins. While historically, researchers have used gels to fractionate proteins, new techniques such as solution-phase, pi-based fractionation, and reversed-phase separations with macroporous LC columns provide high loadability, resolution, and reproducibility, and are easier to automate. These new methods also are enabling the successful fractionation of difficult samples such as membrane proteins, as well as opening up rich possibilities for biomarker discovery. References (1) P.E. Michel, F. Reymond, I.L. Arnaud, J. Josserand, H.H. Girault, and J.S. Rossier, Electrophoresis 24, 3 11 (2003). (2) M. Heller, M. Ye, P.E. Michel, P. Morier, D. Stadler, M.A. Junger, R. Aebersold, F. Reymond, and J.S. Rossier, J. Proteome Res. 4(6), (2005). (3) W. Barrett, J. Martosella, N. Zolotarjova, L-S. Yang, C. Szafranski, and G. Nicol, Agilent publication number EN (2005). (4) W. Barrett, J. Martosella, B. Boyes, C. Szafranski, and G. Nicol, Agilent publication number EN (2005). (5) J. Martosella, N. Zolotarjova, H. Liu, G. Nicol, and B.E. Boyes, J. Proteome Res. 4(5), (2005). (6) N. Zolotarjova, J. Martosella, P. Mrozinski, H. Chen, H. Liu, W. Barrett, and J. Bailey, Multi-Dimensional Workflow Approach for Human Plasma Analysis, Poster 231, HPLC 2006 (2006). (7) J. Martosella, N. Zolotarjova, H. Liu, S. Moyer, P. Perkins, and B.E. Boyes, J. Proteome Res. 5(6), (2006). (8) J. Martosella, N. Zolotarjova, and H. Liu, Agilent publication number EN (2006). (9) A.S. Essader, B.J. Cargile, J.L. Bundy, and J.L. Stephenson Jr., Proteomics 5, (2005). (10) G. Maccarrone, I. Birg, E. Malisch, M.C. Rosenhagen, C. Ditzen, J.A. Chakel, F. Mandel, A. Reimann, C.C. Doertbudak, K. Haegler, F. Holsboer, and C.W. Turck, Clinical Proteomics 1, (2004). (11) M. Heller, P.E. Michel, P. Morier, D. Crettaz, C. Wenz, J-D. Tissot, F. Reymond, and J.S. Rossier, Electrophoresis 26, (2005). (12) C. Miller, S.T. Hoehn, P. Hoerth, and T. Preckel, Enhanced Reproducibility and Predictability in Serum Proteomics Achieved by Off-Gel Isoelectric Focusing of Tryptic Peptides, Poster 132, ABRF 2006 (2006). Jerome Bailey is Program Manager, Bioseparations, Agilent Technologies, Little Falls, Delaware. Peter Mrozinski is an Application Scientist, Bioseparations, Agilent Technologies, Little Falls, Delaware. Tobias Preckel is Product Manager, OFFGEL Fractionator, Agilent Technologies, Waldbronn, Germany. Christine Miller is an Application Scientist, LC MS, Agilent Technologies, Santa Clara, California. James Martosella is an R&D Scientist, Bioseparations, Agilent Technologies, Little Falls, Delaware. Robert Kincaid is a Senior Research Scientist, Agilent Laboratories, Santa Clara, California. Circle 13
Agilent 3100 OFFGEL Fractionator pi-based fractionation of proteins and peptides with liquid-phase recovery
Agilent 3100 OFFGEL Fractionator pi-based fractionation of proteins and peptides with liquid-phase recovery Achieve unprecedented sensitivity in LC/MS-based proteomics experiments Sample complexity and
More informationAlternative to 2D gel electrophoresis OFFGEL electrophoresis combined with high-sensitivity on-chip protein detection.
Alternative to 2D gel electrophoresis OFFGEL electrophoresis combined with high-sensitivity on-chip protein detection Application Note Christian Wenz Andreas Rüfer Abstract Agilent Equipment Agilent 3100
More informationComparative Analysis of Serum Samples with the Agilent mrp-c18 Column - A New Technology for Protein Fractionation Application
Comparative Analysis of Serum Samples with the Agilent mrp-c18 Column - A New Technology for Protein Fractionation Application Proteomics Authors William Barrett, James Martosella, Nina Zolotarjova, Liang-Sheng
More informationIncreasing Sensitivity In LC-MS. Simplification and prefractionation of complex protein samples
Increasing Sensitivity In LC-MS Simplification and prefractionation of complex protein samples Protein Analysis Workflow Simplify Fractionate Separate Identify High Abundant Protein Removal reduction of
More informationConventional isoelectric focusing with the Agilent 3100 OFFGEL Fractionator
Conventional isoelectric focusing with the Agilent 3100 OFFGEL Fractionator Technical Overview Introduction This Technical Overview demonstrates the ability of the Agilent 3100 OFFGEL Fractionator to fractionate
More informationAgilent Multiple Affinity Removal Spin Cartridges for the Depletion of High-Abundant Proteins from Human Proteomic Samples.
Agilent Multiple Affinity Removal Spin Cartridges for the Depletion of High-Abundant Proteins from Human Proteomic Samples Instructions Third edition July 2005 General Information Introduction The Agilent
More informationFaster, easier, flexible proteomics solutions
Agilent HPLC-Chip LC/MS Faster, easier, flexible proteomics solutions Our measure is your success. products applications soft ware services Phospho- Anaysis Intact Glycan & Glycoprotein Agilent s HPLC-Chip
More informationRemoval of Multiple High-Abundant Proteins from Mouse Plasma Using the Agilent Multiple Affinity Removal System for Mouse Application
Removal of Multiple High-Abundant Proteins from Mouse Plasma Using the Agilent Multiple Affinity Removal System for Mouse Application Proteomics Authors William Barrett, Nina Zolotarjova, Haiying Chen,
More informationSeparation of Native Monoclonal Antibodies and Identification of Charge Variants:
Separation of Native Monoclonal Antibodies and Identification of Charge Variants: Teamwork of the Agilent 31 OFFGEL Fractionator, Agilent 21 Bioanalyzer and Agilent LC/MS Systems Application Note Biosimilar
More informationProteome Profiling by Multidimensional Protein Separation and Automated Lab-on-a-Chip Technology Application
Proteome Profiling by Multidimensional Protein Separation and Automated Lab-on-a-Chip Technology Application Proteomics Authors Peter Mrozinski, Bill Wilson, and Hongbin Liu Agilent Technologies, Inc.
More informationKinetics Review. Tonight at 7 PM Phys 204 We will do two problems on the board (additional ones than in the problem sets)
Quiz 1 Kinetics Review Tonight at 7 PM Phys 204 We will do two problems on the board (additional ones than in the problem sets) I will post the problems with solutions on Toolkit for those that can t make
More informationA Highly Accurate Mass Profiling Approach to Protein Biomarker Discovery Using HPLC-Chip/ MS-Enabled ESI-TOF MS
Application Note PROTEOMICS METABOLOMICS GENOMICS INFORMATICS GLYILEVALCYSGLUGLNALASERLEUASPARG CYSVALLYSPROLYSPHETYRTHRLEUHISLYS A Highly Accurate Mass Profiling Approach to Protein Biomarker Discovery
More informationChapter 6. Techniques of Protein and Nucleic Acid Purification
Chapter 6 Techniques of Protein and Nucleic Acid Purification Considerations in protein expression and purification Protein source Natural sources Recombinant sources Methods of lysis and solubilization
More informationMagSi Beads. Magnetic Silica Beads for Life Science and Biotechnology study
MagSi Beads Magnetic Silica Beads for Life Science and Biotechnology study MagnaMedics Diagnostics B.V. / Rev. 9.2 / 2012 Wide range of products for numerous applications MagnaMedics separation solutions
More informationAgilent AssayMAP Platform BIOLOGICS SAMPLE PREPARATION INTELLIGENTLY AUTOMATED
Agilent AssayMAP Platform BIOLOGICS SAMPLE PREPARATION INTELLIGENTLY AUTOMATED SIMPLIFY BIOLOGICS SAMPLE PREPARATION The Agilent AssayMAP platform is an open access, walkaway automation solution specifi
More informationAgilent 3100 OFFGEL Fractionator Kit
Agilent 3100 OFFGEL Fractionator Kit Quick Start Guide Before you begin 2 Reagent Kit Checklist 2 Required Equipment and Reagents 2 Sample and Stock Solution Preparation Notes 3 Procedures 5 To prepare
More informationAgilent AssayMAP Bravo Platform AUTOMATED PROTEIN AND PEPTIDE SAMPLE PREPARATION FOR MASS SPEC ANALYSIS
Agilent AssayMAP Bravo Platform AUTOMATED PROTEIN AND PEPTIDE SAMPLE PREPARATION FOR MASS SPEC ANALYSIS GET FAST, ACCURATE, REPRODUCIBLE RESULTS The Agilent AssayMAP platform is an easy to use yet flexible
More informationAgilent Human 14 Multiple Affinity Removal System Spin Cartridges for the Depletion of High-Abundant Proteins from Human Proteomic Samples
Agilent Human 14 Multiple Affinity Removal System Spin Cartridges for the Depletion of High-Abundant Proteins from Human Proteomic Samples Instructions Second edition October 2008 General Information Introduction
More informationDiscover New Proteins Using Immunodepletion
Discover New Proteins Using Immunodepletion Proteomics Peter Mrozinski Application Scientist Agilent Technologies December 11, 2007 Introduction Complexity and dynamic range of protein concentrations present
More informationSolutions Guide. MX Series II Modular Automation for Nano and Analytical Scale HPLC And Low Pressure Fluid Switching Applications
Solutions Guide MX Series II Modular Automation for Nano and Analytical Scale HPLC And Low Pressure Fluid Switching Applications Page 1 of 12 Table of Contents Sample Injection... 3 Two- Selection... 4
More informationQuantification of Host Cell Protein Impurities Using the Agilent 1290 Infinity II LC Coupled with the 6495B Triple Quadrupole LC/MS System
Application Note Biotherapeutics Quantification of Host Cell Protein Impurities Using the Agilent 9 Infinity II LC Coupled with the 6495B Triple Quadrupole LC/MS System Authors Linfeng Wu and Yanan Yang
More informationSize Exclusion BioHPLC columns Ion Exchange BioHPLC columns
Confidently separate and characterize bio-molecules with Agilent BioHPLC columns Size Exclusion BioHPLC columns Ion Exchange BioHPLC columns "It's a struggle to isolate and identify charge variants of
More informationHichrom Limited 1 The Markham Centre, Station Road, Theale, Reading, Berkshire, RG7 4PE, UK Tel: +44 (0) Fax: +44 (0)
Hichrom Limited 1 The Markham Centre, Station Road, Theale, Reading, Berkshire, RG7 4PE, UK Tel: +44 (0)118 930 3660 Fax: +44 (0)118 932 3484 Email: www.hichrom.co.uk Table of Contents Sample Injection...
More informationProteomics. Proteomics is the study of all proteins within organism. Challenges
Proteomics Proteomics is the study of all proteins within organism. Challenges 1. The proteome is larger than the genome due to alternative splicing and protein modification. As we have said before we
More informationProtein analysis. Dr. Mamoun Ahram Summer semester, Resources This lecture Campbell and Farrell s Biochemistry, Chapters 5
Protein analysis Dr. Mamoun Ahram Summer semester, 2015-2016 Resources This lecture Campbell and Farrell s Biochemistry, Chapters 5 Bases of protein separation Proteins can be purified on the basis Solubility
More informationFast and Efficient Peptide Mapping of a Monoclonal Antibody (mab): UHPLC Performance with Superficially Porous Particles
Fast and Efficient Peptide Mapping of a Monoclonal Antibody (mab): UHPLC Performance with Superficially Porous Particles Application Note Biotherapeutics and Biosimilars Authors James Martosella, Alex
More informationIdentification of human serum proteins detectable after Albumin removal with Vivapure Anti-HSA Kit
Andreas Kocourek, Pieter Eyckerman, Robert Zeidler, Pascal Bolon and Birgit Thome-Kromer Introduction The analysis of the complete proteome is a major interest of many researchers. Of particular importance
More informationFOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE.
User Protocol 122643 Rev. 12 May 2005 JSW Page 1 of 7 ProteoExtract Albumin/IgG Removal Kit, Maxi Cat. No. 122643 1. Introduction One of the major challenges in functional proteomics is the handling of
More informationNPTEL VIDEO COURSE PROTEOMICS PROF. SANJEEVA SRIVASTAVA
LECTURE-06 PROTEIN PURIFICATION AND PEPTIDE ISOLATION USING CHROMATOGRAPHY TRANSCRIPT Welcome to the proteomics course. Today, we will talk about protein purification and peptide isolation using chromatography
More informationMass Spectrometry Analysis of Liquid Chromatography Fractions using Ettan LC MS System
GUIDE TO LC MS - December 21 1 Spectrometry Analysis of Liquid Chromatography Fractions using Ettan LC MS System Henrik Wadensten, Inger Salomonsson, Staffan Lindqvist, Staffan Renlund, Amersham Biosciences,
More informationWhite Paper. Ion Exchange with PureSpeed Tips A Powerful Chromatography Tool
Ion Exchange with PureSpeed Tips A Powerful Chromatography Tool Ion exchange chromatography separates molecules by exploiting differences in their overall charge characteristics. Its simplicity makes this
More informationAgilent 6430 Triple Quadrupole LC/MS System
Agilent 6430 Triple Quadrupole LC/MS System Ideal Quantitative LC/MS for UHPLC with Dynamic MRM and Fast Polarity Switching, plus Unsurpassed Sensitivity with Chip LC Summary The Agilent 6430 Triple Quadrupole
More informationProteomics and some of its Mass Spectrometric Applications
Proteomics and some of its Mass Spectrometric Applications What? Large scale screening of proteins, their expression, modifications and interactions by using high-throughput approaches 2 1 Why? The number
More informationcolumns P r o P a c H I C C o l u m n S o l u t i o n s f o r P r o t e i n A n a l y s i s
columns P r o P a c H I C - 1 C o l u m n S o l u t i o n s f o r P r o t e i n A n a l y s i s Hydrophobic Interaction Chromatography (HIC) is an important tool for protein chemists separating proteins
More informationLaboratory Water Quality Affects Protein Separation by 2D Gel Electrophoresis
Laboratory Water Quality Affects Protein Separation by 2D Gel Electrophoresis 2D gel electrophoresis remains a dominant technique in proteomics. Obtaining high quality gels requires careful and tedious
More informationColumn for High Performance,High-Binding Capacity Ion Exchange Chromatography:TSKgel SuperQ-5PW and Its Applications
ANALYSIS S e p a r a t i o n R e p o r t N o. 9 3 Column for High Performance,High-Binding Capacity Ion Exchange Chromatography:TSKgel SuperQ-5PW and Its Applications Table of Contents 1. Introduction
More informationStrategies in proteomics
Strategies in proteomics Systems biology - understand cellpathways, network, and complex interacting (includes Genomics, Proteomics, Metabolomics..) Biological processes - characterize protein complexes,
More informationPLRP-S Polymeric Reversed-Phase Column for LC/MS Separation of mabs and ADC
PLRP-S Polymeric Reversed-Phase Column for LC/MS Separation of mabs and ADC Analysis of Intact and Fragmented mabs and ADC Application Note Biotherapeutics and Biologics Author Suresh Babu C.V. Agilent
More informationYour complete protein clean-up toolbox
Your complete protein clean-up toolbox Thermo Scientific protein sample clean-up devices have evolved over the last decade to meet a variety of customer needs. Our broad portfolio of high-performance tools
More informationSeparation of Monoclonal Antibodies Using TSKgel HPLC Columns
ANALYSIS S e p a r a t i o n R e p o r t N o. 7 4 Separation of Monoclonal Antibodies Using TSKgel HPLC Columns Table of Contents 1. Introduction 1 2. Separation Mode and Purification Method 1 3. Applications
More informationspecial offers from your protein biology resource
special offers from your protein biology resource Pop open your cells, extract your proteins, purify, quantify and express them. Seeking knowledge about proteins with Thermo Scientific Protein Research
More informationAccelerate mab Characterization Using Automated Sample Prep
Accelerate mab Characterization Using Automated Sample Prep David Knorr, Ph.D. Automation Solutions Ning Tang, Ph.D. LC/MS 15 February 2012 Page 1 Protein Sample Processing Workflows Glycan Profiling Biological
More informationProteomics and Cancer
Proteomics and Cancer Japan Society for the Promotion of Science (JSPS) Science Dialogue Program at Niitsu Senior High School Niitsu, Niigata September 4th 2006 Vladimir Valera, M.D, PhD JSPS Postdoctoral
More informationLecture 5: 8/31. CHAPTER 5 Techniques in Protein Biochemistry
Lecture 5: 8/31 CHAPTER 5 Techniques in Protein Biochemistry Chapter 5 Outline The proteome is the entire set of proteins expressed and modified by a cell under a particular set of biochemical conditions.
More informationIsolation of Protein
Isolation of Protein Ultra-centrifugation http://irfanchemist.wordpress.com/2009/04/19/isolation-of-protein / Protein solutions of various masses or densities may separated based on the time it takes to
More information12/6/12. Dr. Sanjeeva Srivastava IIT Bombay
Dr. Sanjeeva Srivastava IIT Bombay Gel-based proteomics 2-DE work-flow 1 Isoelectric focusing (first dimension) 2 Equilibration of IPG strips 3 SDS-PAGE (second dimension) 4 Staining gel visualization
More informationS R E VA V Blu l e P ep Kit i s SERVA Blue Prep CBD Micro/MacroKit SERVA Blue Prep DetergentEx Micro/Macro Kit SERVA Blue
Kits Protein and Protein/Nucleic acid Isolation and Purification Kits SERVA BluePrep CBD Micro/MacroKit SERVA BluePrep DetergentEx Micro/Macro Kit SERVA BluePrep IB Isolation Micro/Macro Kit Cell Lysis
More informationApplication Note 18 RNA/DNA/Protein Sample Preparation METHODS AND MATERIALS INTRODUCTION
Application Note 18 /DNA/Protein Sample Preparation Sequential Purification of, DNA and Protein from a Single Sample using 's /DNA/Protein Purification Kit and Comparison to a Market B. Lam, PhD 1, C.
More informationMass Spectrometry and Proteomics - Lecture 6 - Matthias Trost Newcastle University
Mass Spectrometry and Proteomics - Lecture 6 - Matthias Trost Newcastle University matthias.trost@ncl.ac.uk Previously Quantitation techniques Label-free TMT SILAC Peptide identification and FDR 194 Lecture
More informationMETHODS IN CELL BIOLOGY EXAM II, MARCH 26, 2008
NAME KEY METHODS IN CELL BIOLOGY EXAM II, MARCH 26, 2008 1. DEFINITIONS (30 points). Briefly (1-3 sentences, phrases, word, etc.) define the following terms or answer question. A. depot effect refers to
More informationPeptide enrichment and fractionation
Protein sample preparation Successful analysis of low-abundance proteins and/or the identification of posttranslationally modified peptides often require several steps: enrichment, fractionation, and/or
More informationApplication Manual ProteoSpin Abundant Serum Protein Depletion Kit
Application Manual ProteoSpin Abundant Serum Protein Depletion Kit For Use with P/N 17300 ProteoSpin Abundant Serum Protein Depletion Kit Basic Features Efficient removal of highly abundant proteins 1
More informationRapid Extraction of Therapeutic Oligonucleotides from Primary Tissues for LC/ MS Analysis Using Clarity OTX, an Oligonucleotide Extraction Cartridge
Rapid Extraction of Therapeutic Oligonucleotides from Primary Tissues for LC/ MS Analysis Using Clarity OTX, an Oligonucleotide Extraction Cartridge G. Scott*, H. Gaus #, B. Rivera*, and M. McGinley* *Phenomenex,
More informationProSEC 300S. Protein Characterization columns
ProSEC 300S Protein Characterization columns Agilent s ProSEC 300S is a silica-based material specifically designed for the analysis of proteins by aqueous size exclusion chromatography. With a proprietary
More informationApplication Note # ET-20 BioPharma Compass: A fully Automated Solution for Characterization and QC of Intact and Digested Proteins
Application Note # ET-20 BioPharma Compass: A fully Automated Solution for Characterization and QC of Intact and Digested Proteins BioPharma Compass TM is a fully automated solution for the rapid characterization
More information(Refer Slide Time: 00:16)
(Refer Slide Time: 00:16) Proteins and Gel-Based Proteomics Professor Sanjeeva Srivastava Department of Biosciences and Bioengineering Indian Institute of Technology, Bombay Mod 02 Lecture Number 5 Welcome
More informationGELFREE 8100 Fractionation System. Frequently Asked Questions
INDEX Basics...3 Features...3 Sample Loading...4 Operation...5 Fraction Recovery...6 Tips...7 Troubleshooting...7 Use and Use Restrictions: The Products are sold, and deliverables of any services are provided
More informationProteome Purify TM 12
Proteome Purify TM 12 Human Serum Protein Immunodepletion Resin Catalog Number IDR012-020 Catalog Number IDR012-040 For the removal of twelve high-abundance proteins from human serum or plasma. This package
More informationJet Stream Proteomics for Sensitive and Robust Standard Flow LC/MS
Jet Stream Proteomics for Sensitive and Robust Standard Flow LC/MS Technical Overview Authors Yanan Yang, Vadiraj hat, and Christine Miller Agilent Technologies, Inc. Santa Clara, California Introduction
More information2 Liquid chromatography of biomolecules
2 Liquid chromatography of biomolecules Proteins, peptides, DNA, RNA, lipids, and organic cofactors have various characteristics such as electric charge, molecular weight, hydrophobicity, and surface relief.
More informationAn effective platform for purification of IgM monoclonal antibodies using Hydroxyapatite
An effective platform for purification of IgM monoclonal antibodies using Hydroxyapatite Frank Hensel, Patrys, GmbH Pete Gagnon, Validated Biosystems 5th International Conference on Hydroxyapatite and
More informationAgilent Prep LC Columns for Small Molecules and Biomolecules MAINTAIN RAPID, RELIABLE SEPARATIONS AS YOU SCALE-UP
Agilent Prep LC Columns for Small Molecules and Biomolecules MAINTAIN RAPID, RELIABLE SEPARATIONS AS YOU SCALE-UP AGILENT PREP COLUMNS FOR HPLC FLEXIBLE, COST-EFFECTIVE OPTIONS FOR SCALING AND PREPARATIVE
More informationALP (alkaline phosphatase) calibrators were analyzed manually in microtiter plates to find the linearity range by following this protocol:
Exam Mol 3008 May 2009 Subject 1 (15p) ALP (alkaline phosphatase) calibrators were analyzed manually in microtiter plates to find the linearity range by following this protocol: Reaction solutions: 50
More information11/22/13. Proteomics, functional genomics, and systems biology. Biosciences 741: Genomics Fall, 2013 Week 11
Proteomics, functional genomics, and systems biology Biosciences 741: Genomics Fall, 2013 Week 11 1 Figure 6.1 The future of genomics Functional Genomics The field of functional genomics represents the
More informationsample preparation Accessing Low-Abundance Proteins in Serum and Plasma With a Novel, Simple Enrichment and Depletion Method tech note 5632
sample preparation tech note 5632 Accessing Low-Abundance Proteins in Serum and Plasma With a Novel, Simple Enrichment and Depletion Method Aran Paulus, Steve Freeby, Katrina Academia, Vanitha Thulasiraman,
More informationProteoExtract Protein Precipitation Kit Cat. No
User Protocol 539180 Rev. 12-October-06 JSW ProteoExtract Protein Precipitation Kit Cat. No. 539180 Note that this user protocol is not lot-specific and is representative of the current specifications
More informationReducing dynamic range with Proteominer
Reducing dynamic range with Proteominer Dynamic Range of the Plasma Proteome Albumin Apolipo-A1 Apolipo-B AGP Lipoprotein A IgG Transferrin Fibrinogen IgA α2 macroglobulin IgM α1-at C3 Comp factor Haptoglobulin
More information1. Bloomsbury BBSRC Centre for Structural Biology, Birkbeck College and University College London.
Purification/Polishing of His-tagged proteins - Application of Centrifugal Vivapure Ion-exchange Membrane Devices to the Purification/Polishing of Histagged Background Multi-milligram quantities of highly
More informationProteomics And Cancer Biomarker Discovery. Dr. Zahid Khan Institute of chemical Sciences (ICS) University of Peshawar. Overview. Cancer.
Proteomics And Cancer Biomarker Discovery Dr. Zahid Khan Institute of chemical Sciences (ICS) University of Peshawar Overview Proteomics Cancer Aims Tools Data Base search Challenges Summary 1 Overview
More informationProteoEnrich CAT-X SEC Kit Merck: Lichrospher Composition. Silica Particle size. 25 µm particles with 6 nm pore size Ligand SO 3
Novagen User Protocol TB428 Rev. A 1105 1 of 6 ProteoEnrich TM CAT-X SEC Kit About the Kit ProteoEnrich CAT-X SEC Kit 71539-3 Description The ProteoEnrich CAT-X SEC Kit provides a highly specific method
More informationSimplifying ImmunoAffinity Capture Workflow
CAE TUDY MART Digest ImmunoAffinity (IA) Kit implifying ImmunoAffinity Capture Workflow Rapid, ensitive, LC-RM Quantitative Analysis of Proteins in Plasma As pharmaceuticals grow more efficacious, reporting
More informationTSK-GEL BioAssist Series Ion Exchange Columns
Separation Report No. 100 TSK-GEL BioAssist Series Ion Exchange Columns Table of Contents 1. Introduction 2 2. Basic Properties 2 2-1 Ion-Exchange Capacity and Pore Characteristics 2 2-2 Separation of
More informationWhy purify proteins?
Why purify proteins? Detailed studies on function Determination of structure Industrial/pharmaceutical applications Generate antibodies Amino acid sequence determination 1/16/04 Marilyn Niemann, UAB/CORD
More informationProteomics Background and clinical utility
Proteomics Background and clinical utility H.H. Helgason MD Antoni van Leeuwenhoek Hospital The Netherlands Cancer Institute Amsterdam Introduction Background Definitions Protein biomarkers Technical aspects
More informationMaximizing Chromatographic Resolution of Peptide Maps using UPLC with Tandem Columns
Maximizing Chromatographic Resolution of Peptide Maps using UPLC with Tandem Columns Hongwei Xie, Martin Gilar, and Jeff Mazzeo Waters Corporation, Milford, MA U.S. APPLICATION BENEFITS The ACQUITY UPLC
More informationLecture 7: Affinity Chromatography-II
Lecture 7: Affinity Chromatography-II We have studied basics of affinity purification during last lecture. The current lecture is continuation of last lecture and we will cover following: 1. Few specific
More informationsample preparation Accessing Low-Abundance Proteins in Serum and Plasma With a Novel, Simple Enrichment and Depletion Method tech note 5632
sample preparation tech note 5632 Accessing Low-Abundance Proteins in Serum and Plasma With a Novel, Simple Enrichment and Depletion Method Aran Paulus, Steve Freeby, Katrina Academia, Vanitha Thulasiraman,
More informationPROCEDURE FOR USE NICKEL NTA Magnetic Agarose Beads (5%)
1 AFFINITY HIS-TAG PURIFICATION PROCEDURE FOR USE NICKEL NTA Magnetic Agarose Beads (5%) DESCRIPTION Nickel NTA Magnetic Agarose Beads are products that allow rapid and easy small-scale purification of
More informationMonoclonal Antibody Analysis on a Reversed-Phase C4 Polymer Monolith Column
Monoclonal Antibody Analysis on a Reversed-Phase C4 Polymer Monolith Column Shane Bechler 1, Ken Cook 2, and Kelly Flook 1 1 Thermo Fisher Scientific, Sunnyvale, CA, USA; 2 Thermo Fisher Scientific, Runcorn,
More informationApplication Note. Authors. Abstract. Ravindra Gudihal Agilent Technologies India Pvt. Ltd. Bangalore India
Characterization of bacteriophage derived anti-staphylococcal protein (P28) from production to purification using Agilent HPLC-Chip Q-TOF LC/MS system Application Note Authors Ravindra Gudihal Agilent
More informationChromatography column for therapeutic protein analysis
PRODUCT SPECIFICATIONS ProPac Elite WCX Column Chromatography column for therapeutic protein analysis Benefits Superior resolution power for proteins, monoclonal antibodies, and associated charge variants
More informationComparison of different methods for purification analysis of a green fluorescent Strep-tag fusion protein. Application
Comparison of different methods for purification analysis of a green fluorescent Strep-tag fusion protein Application Petra Sebastian Meike Kuschel Stefan Schmidt Abstract This Application Note describes
More informationProteoMiner Protein Enrichment Technology
Sample Preparation ProteoMiner Protein Enrichment Technology Digging Deeper in the Proteome Detect More Proteins With ProteoMiner Technology Than With Immunodepletion ProteoMiner Technology vs. an Agilent
More informationHigh-resolution Analysis of Charge Heterogeneity in Monoclonal Antibodies Using ph-gradient Cation Exchange Chromatography
High-resolution Analysis of Charge Heterogeneity in Monoclonal Antibodies Using ph-gradient Cation Exchange Chromatography Agilent 1260 Infinity Bio-inert Quaternary LC System with Agilent Bio Columns
More informationExtracting Pure Proteins from Cells
Extracting Pure Proteins from Cells 0 Purification techniques focus mainly on size & charge 0 The first step is homogenization (grinding, Potter Elvejhem homogenizer, sonication, freezing and thawing,
More informationLecture 8: Affinity Chromatography-III
Lecture 8: Affinity Chromatography-III Key words: Chromatography; Affinity chromatography; Protein Purification During this lecture, we shall be studying few more examples of affinity chromatography. The
More informationComparison of Different Brands of Carrier Ampholytes for Monoclonal Antibody Charge Heterogeneity Analysis by Capillary Isoelectric Focusing
Comparison of Different Brands of Carrier Ampholytes for Monoclonal Antibody Charge Heterogeneity Analysis by Capillary Isoelectric Focusing Application Note Biotherapeutics & Biosimilars Author Christian
More informationProteoSpin Urine Protein Concentration Midi Kit Product # 52300
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com ProteoSpin Urine Protein Concentration Midi Kit Product # 52300
More informationProteoSpin Urine Protein Concentration Maxi Kit Product # 21600
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com ProteoSpin Urine Protein Concentration Maxi Kit Product # 21600
More informationFor the quick and efficient purification of highly specific and ultra pure antibodies
ab138915 EpiMAX Affinity Purification Kit Instructions for Use For the quick and efficient purification of highly specific and ultra pure antibodies This product is for research use only and is not intended
More informationSeparation of Recombinant Human Erythropoietin (repo) Using Agilent Bio SEC-3
Separation of Recombinant Human Erythropoietin (repo) Using Agilent Bio SEC-3 Application Note BioPharma Authors Phu T Duong and James Martosella Agilent Technologies, Inc. 285 Centerville Rd, Wilmington,
More informationPeptide Mapping: A Quality by Design (QbD) Approach
Peptide Mapping: A Quality by Design (QbD) Approach Application Note Bio-Pharmaceutical Authors Sreelakshmy Menon and Suresh babu C.V. Agilent Technologies, Inc. Richard Verseput S-Matrix Corporation Abstract
More informationAgilent SD-1 Purification System. Purify your way SD-1
Agilent SD-1 Purification System Purify your way SD-1 AGILENT SD-1 PURIFICATION SYSTEM PURIFY YOUR WAY WITH HIGH QUALITY SEPARATIONS AT ANY SCALE The Agilent SD-1 Purification System achieves better gradient
More informationPureSpeed Tips. Superior Protein Purity and Concentration Protein Purification in a Pipette Tip
PureSpeed Tips PureSpeed Protein Tips Highest purity and concentration Fast as little as 15 minutes Process many samples at once Superior Protein Purity and Concentration Protein Purification in a Pipette
More informationAdvanceBio HIC: a Hydrophobic HPLC Column for Monoclonal Antibody (mab) Variant Analysis
Application Note Biologics Development AdvanceBio HIC: a Hydrophobic HPLC Column for Monoclonal Antibody (mab) Variant Analysis Using the Agilent 16 Infinity II Bio-Inert LC Authors Andrew Coffey and Sandeep
More informationIntroduction to Proteomics
Introduction to Proteomics Tasso Miliotis, PhD AstraZeneca R&D Gothenburg Translational Sciences tasso.miliotis@astrazeneca.com 1 Site Management Mölndal May 2008 Outline Drug Discovery AZ R&D Sample Preparation
More informationDISCOVERY AND VALIDATION OF TARGETS AND BIOMARKERS BY MASS SPECTROMETRY-BASED PROTEOMICS. September, 2011
DISCOVERY AND VALIDATION OF TARGETS AND BIOMARKERS BY MASS SPECTROMETRY-BASED PROTEOMICS September, 2011 1 CAPRION PROTEOMICS Leading proteomics-based service provider - Biomarker and target discovery
More informationProtein Methods. Second Edition. DANIEL M. BOLLAG Merck Research Laboratories West Point, Pennsylvania
Protein Methods Second Edition DANIEL M. BOLLAG Merck Research Laboratories West Point, Pennsylvania MICHAEL D. ROZYCKI Department of Chemistry The Henry H. Hoyt Laboratory Princeton University Princeton,
More informationDownstream Processing (DSP): Microscale Automation of Biopharmaceutical Processes. Michel Eppink, Synthon BV, Nijmegen, The Netherlands
Downstream Processing (DSP): Microscale Automation of Biopharmaceutical Processes Michel Eppink, Synthon BV, Nijmegen, The Netherlands Overview Introduction Why automation? Automation in DSP processes
More informationAgilent 1290 Infinity II 2D-LC Solution Biopharmaceutical Polymer Analysis. WCBP Jan 2017 Washington, DC
Agilent 1290 Infinity II 2D-LC Solution Biopharmaceutical Polymer Analysis WCBP Jan 2017 Washington, DC 1 Overview Resolving power and how to measure it Why two-dimensional LC? Setup of a 2D-LC System
More information