This document summarizes a study comparing the use of electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) for liquid chromatography-mass spectrometry (LC-MS) workflows. The study found that ESI generally provides higher peptide sequence coverage than MALDI. MALDI detection is better correlated with larger, more hydrophobic peptides having lower isoelectric points, while ESI detection favors smaller peptides with higher isoelectric points. The optimal choice of ionization method depends on the application and type of samples being analyzed.
Proteomics is the study of the structure and function of proteins. It involves identifying and quantifying the proteins expressed by a genome or cell type. Key aspects of proteomics include protein separation techniques like gel electrophoresis, mass spectrometry to identify proteins, and analyzing protein interactions and post-translational modifications. While genomes provide the blueprint, proteomics helps understand the diversity of proteins expressed and how they function together to direct cellular activities. It is a promising tool for disease diagnosis by identifying protein biomarkers.
BITS - Introduction to Mass Spec data generationBITS
Mass spectrometry is a technique that analyzes molecules based on their mass-to-charge ratio. It involves ionizing samples using sources like MALDI or ESI, separating the ions using mass analyzers like time-of-flight or quadrupole, and detecting the ions. Tandem mass spectrometry allows targeted fragmentation of selected ions to gain additional structural information. Amino acids are the building blocks of proteins and contain variable side chains attached to a common peptide backbone. Mass spectrometry is useful for analyzing proteins and peptides.
Gene cloning involves isolating a particular gene or DNA fragment of interest from an organism's total DNA and producing many copies of just that fragment. There are several reasons for cloning DNA, such as determining a gene's nucleotide sequence, identifying control sequences, investigating protein/enzyme function, and engineering organisms for specific purposes like insulin production. Common tools used in cloning include restriction enzymes, ligase, vectors like plasmids and bacteriophages, and host cells. DNA is cut with restriction enzymes, ligated into a vector, and introduced into host cells to replicate the exogenous DNA fragment.
The document describes the development of an enhanced performance test mix for monitoring high-throughput LC/MS systems used in pharmaceutical analysis. The test mix was designed to:
1) Consist of 8 drug-like compounds selected from a pool of 137 compounds to represent typical properties like mass, hydrophobicity, and charge state.
2) Monitor key aspects of LC/MS performance including separation (gradient, flow, pH), detection (UV, ELS, MS signal), and mass accuracy across different conditions.
3) Provide diagnostic information about the likely cause of any errors based on differences observed in test mix results compared to historical data.
This document summarizes a study comparing traditional immunoassay screening followed by confirmation using solid phase extraction and gas chromatography/mass spectrometry (SPE/GC-MS) or liquid chromatography/tandem mass spectrometry (LC-MS/MS) to a new method using automated solid phase extraction coupled to high performance liquid chromatography and tandem mass spectrometry (SPE/HPLC/MS/MS) for analyzing urine samples submitted for driving under the influence (DUI) cases. In the study, 106 samples previously confirmed positive by the traditional methods were re-analyzed using the new automated SPE/HPLC/MS/MS method. The new method found additional positive results for THC, amphetamines, benzodiazep
Proteomics is the study of the structure and function of proteins. It involves identifying and quantifying the proteins expressed by a genome or cell type. Key aspects of proteomics include protein separation techniques like gel electrophoresis, mass spectrometry to identify proteins, and analyzing protein interactions and post-translational modifications. While genomes provide the blueprint, proteomics helps understand the diversity of proteins expressed and how they function together to direct cellular activities. It is a promising tool for disease diagnosis by identifying protein biomarkers.
BITS - Introduction to Mass Spec data generationBITS
Mass spectrometry is a technique that analyzes molecules based on their mass-to-charge ratio. It involves ionizing samples using sources like MALDI or ESI, separating the ions using mass analyzers like time-of-flight or quadrupole, and detecting the ions. Tandem mass spectrometry allows targeted fragmentation of selected ions to gain additional structural information. Amino acids are the building blocks of proteins and contain variable side chains attached to a common peptide backbone. Mass spectrometry is useful for analyzing proteins and peptides.
Gene cloning involves isolating a particular gene or DNA fragment of interest from an organism's total DNA and producing many copies of just that fragment. There are several reasons for cloning DNA, such as determining a gene's nucleotide sequence, identifying control sequences, investigating protein/enzyme function, and engineering organisms for specific purposes like insulin production. Common tools used in cloning include restriction enzymes, ligase, vectors like plasmids and bacteriophages, and host cells. DNA is cut with restriction enzymes, ligated into a vector, and introduced into host cells to replicate the exogenous DNA fragment.
The document describes the development of an enhanced performance test mix for monitoring high-throughput LC/MS systems used in pharmaceutical analysis. The test mix was designed to:
1) Consist of 8 drug-like compounds selected from a pool of 137 compounds to represent typical properties like mass, hydrophobicity, and charge state.
2) Monitor key aspects of LC/MS performance including separation (gradient, flow, pH), detection (UV, ELS, MS signal), and mass accuracy across different conditions.
3) Provide diagnostic information about the likely cause of any errors based on differences observed in test mix results compared to historical data.
This document summarizes a study comparing traditional immunoassay screening followed by confirmation using solid phase extraction and gas chromatography/mass spectrometry (SPE/GC-MS) or liquid chromatography/tandem mass spectrometry (LC-MS/MS) to a new method using automated solid phase extraction coupled to high performance liquid chromatography and tandem mass spectrometry (SPE/HPLC/MS/MS) for analyzing urine samples submitted for driving under the influence (DUI) cases. In the study, 106 samples previously confirmed positive by the traditional methods were re-analyzed using the new automated SPE/HPLC/MS/MS method. The new method found additional positive results for THC, amphetamines, benzodiazep
This document summarizes a study comparing traditional immunoassay screening followed by confirmation using solid phase extraction and gas chromatography/mass spectrometry (SPE/GC-MS) versus automated solid phase extraction high performance liquid chromatography tandem mass spectrometry (SPE/HPLC/MS/MS) for analyzing urine samples submitted for driving under the influence (DUI) cases. 106 samples previously confirmed positive by the traditional methods were re-analyzed using the automated SPE/HPLC/MS/MS system. More drugs were detected using the automated system, including 12 additional THC cases, 4 additional amphetamine cases, 6 additional benzodiazepine cases, and 3 additional cocaine cases. The automated system was able to process samples faster and at
The common labels of chemiluminescent immunoassays is alkaline phosphatase (ALP). Huateng Pharma can provides APS-5(CAS#193884-53-6) from milligram to kilogram scale.
Theoretical evaluation of shotgun proteomic analysis strategies; Peptide obse...Keiji Takamoto
This document discusses evaluating different strategies for shotgun proteomic analysis through theoretical modeling. It develops a peptide observability function based on mouse proteomic data to predict how observable peptides are by LC-MS/MS. This function is applied to theoretically digested mouse proteins using different proteases and separation techniques to evaluate their combinations and the separation profiles achieved. The results suggest SAX/trypsin and IEF/trypsin are favorable combinations that provide good separation.
This document discusses liquid chromatography-mass spectrometry (LC-MS). It begins with an introduction to LC-MS, explaining that it combines liquid chromatography separation with mass spectrometry detection. It then covers the history, principles, instrumentation, advantages, and applications of LC and MS. Key applications discussed include pharmacokinetics, proteomics, metabolomics, and drug development.
This document describes a study comparing data acquired from data-independent LC-MS to data acquired from data-dependent LC-MS/MS. The study analyzed mixtures of four proteins alone and with a complex E. coli protein digest. Each sample was run in triplicate by both acquisition methods. The data-independent LC-MS provided more comprehensive detection of precursor and product ions than the combined data-dependent LC-MS/MS experiments. Over 90% of masses detected by LC-MS/MS were also detected by data-independent LC-MS at the correct retention times with similar fragmentation patterns. The data-independent LC-MS was able to detect more components than the individual data-dependent LC-MS/MS experiments.
Bioanalytical Method Development and Validation for Simultaneous Estimation o...BRNSSPublicationHubI
The document describes the development and validation of a bioanalytical method for the simultaneous estimation of imatinib and its metabolite desmethyl imatinib in human plasma using liquid chromatography-mass spectrometry. Key steps in the method included online enrichment of the analytes followed by separation on a chromatographic column and detection using mass spectrometry. The method was validated in terms of precision, accuracy, selectivity and sensitivity. The developed method was then applied to pharmacokinetic studies of imatinib and its metabolite in patient samples.
ICP-MS detects elements instead of molecules. With the exception of a few elements (C, H, N, O and the noble gases), all elements can be detected. A specific element serves as a tag for the drug molecule of interest, thus enabling quantitation of this drug molecule in a particular matrix. The technique is highly linear and can be used quantitatively for a broad concentration range. Sample processing is relatively easy and throughput times are short, resulting in fast turnaround times.
Liquid chromatography-mass spectrometry (LC-MS) is an analytical technique that combines liquid chromatography with mass spectrometry. It has high sensitivity and selectivity. LC-MS is commonly used in pharmacokinetics studies to analyze drugs in biological samples. It is also used in proteomics to identify over 1000 proteins in complex samples like human serum. Additionally, LC-MS has various applications in drug development such as metabolite identification, quantitative bioanalysis, and quality control.
Proteomics_Chapter 3 Protein Identification.pptZaldaaZaldaa
The document provides information on learning outcomes and an overview of protein identification methods. The key learning outcomes are to understand the workflow of protein identification, identification by Western blotting and mass spectrometry, components of a mass spectrometer, and different ionization and mass spectrometry methods. It then describes in more detail the processes of protein transfer and antibody detection in Western blotting, as well as common ionization techniques like MALDI and ESI and mass analyzer types for protein identification by mass spectrometry.
Adsorptive stripping differential pulse voltammetry determination of rivastig...Pramod Kalambate
The study of graphene nanosheet (GNS)–gold nanoparticle (AuNP)–carbon paste electrode (GNS–AuNP–CPE) as
an electrochemical sensor for the determination of rivastigmine (RIV) in pharmaceuticals formulations, blood
serum, and urine samples is presented. The GNS–AuNP composite is prepared by in situ simultaneous reduction
of graphene oxide and chloroauric acid using sodiumborohydride as a reducing agent. The GNS–AuNP composite
was characterized by X-ray diffraction, UV–Vis spectroscopy, and scanning electron microscopy. Electrochemical
characterization of the GNS–AuNP–CPE electrode surfacewas carried out by cyclic voltammetry, electrochemical
impedance spectroscopy, chronocoulometry, and adsorptive stripping differential pulse voltammetry. This study
shows that oxidation of rivastigmine is facilitated at the GNS–AuNP–CPE electrode and remarkably increase in
current compared to the bare electrode due to enhanced adsorption of the former on electrode surface. Under
the optimized conditions, the peak current (Ip) is found to be proportional to the RIV concentration in the
range of 2.0 × 10–7–6.0 × 10−4 M with a detection limit of 5.3 × 10−8 M. The proposed sensor shows a very
high level of sensitivity, selectivity, and a very good reproducibility for RIV determination. A good recovery
level obtained for real samples suggests practical utility of the GNS–AuNP–CPE as an effective and reliable electrochemical
sensor for RIV detection.
The document provides an introduction to LC-MS, including:
- LC-MS combines liquid chromatography with mass spectrometry to separate and identify components in a sample.
- It can be used for qualitative analysis to determine what is in a sample and quantitative analysis to determine concentrations.
- The document reviews the basic components and workflow of LC-MS, including how the liquid chromatography separates components and the mass spectrometer ionizes and detects them.
- It provides guidance on starting up and running samples on an Agilent 6100 single quadrupole LC-MS system, including loading methods, running samples, and viewing resultant data.
Expanding Surface Plasmon Resonance Capabilities with ReichertReichertSPR
Surface Plasmon Resonance (SPR) is a widely-used label-free technique to characterize a variety of molecular interactions. SPR is an optical phenomenon that is sensitive to changes in the dielectric properties of the medium close to a metal surface. Specifically, the resonance condition is affected by changes in refractive index occurring up to 300 nm above the metal surface (Au) and thus by the material absorbed onto the metal film. Therefore, the SPR signal is a measure of the total mass concentration at the gold sensor chip surface.
Typically, a mobile molecule (analyte) is injected across an immobilized binding partner (ligand) and as the analyte binds, this mass accumulation on the sensor surface leads to an increase in refractive index, and the result is plotted as response versus time. SPR is commonly utilized by researchers to determine association/dissociation rates, affinities and thermodynamics of biomolecular interactions.
Traditionally, the interactions under study with SPR include those occurring with and between the major classes of biological macromolecules along with those involving small molecules and drugs. These classic experiments have been primarily carried with just purified samples. Reichert’s SPR systems implement a very robust fluidics arrangement that can accommodate a wide variety of sample compositions including crude samples such as lysates, whole cells and serum. In addition, Reichert’s systems are housed in an open architecture that easily allows coupling to other analytical techniques and instruments. Along with excelling at traditional biomolecular interactions, Reichert’s systems pave the way for new avenues of investigation involving crude samples and whole cells along with the ability to couple SPR to other techniques. This presentation will focus on the SPR technique and provide examples of unique applications with cells along with the possibility of interfacing SPR with other analytical methods.
http://www.reichertspr.com/webinars/general/expanding-surface-plasmon-resonance-capabilities-with-reichert/
ACCULAB Corporation is a manufacturer and supplier of laboratory and medical instruments based in New York. It incorporates 7 companies to offer a wide range of products at different price points to meet customer demands. ACCULAB aims to provide both high quality products and competitive prices with a vision of being a one-stop shop for all laboratory equipment needs through excellent customer service.
MALDI-TOF: Pricinple and Its Application in Biochemistry and BiotechnologyDevakumar Jain
The document discusses MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry. It provides a history of the development of mass spectrometry techniques. MALDI-TOF allows for the analysis of intact biomolecules like proteins and is a soft ionization method. It provides high sensitivity and mass accuracy for analyzing proteins, peptides, and other large biomolecules. The document also discusses applications of MALDI-TOF like protein identification and characterization.
This document describes a novel database search algorithm for identifying proteins from data independent acquisitions where multiple precursor ions are fragmented simultaneously. The algorithm uses an iterative process to incrementally increase selectivity, specificity, and sensitivity. It accounts for peptide retention time, ion intensities, charge states, and accurate masses of precursors and products. The algorithm was tested on simple and complex protein mixtures and validated independently, demonstrating its ability to correctly identify proteins across a wide dynamic range with high sensitivity and specificity.
Saravanan Kumar,V.Siva Reddy - Workshop on ProteomicsSARAVANAN KUMAR
1) Mass spectrometry is a technique that measures the mass-to-charge ratio of ions based on how they move in electric or magnetic fields.
2) Samples are ionized using techniques like electrospray ionization or matrix-assisted laser desorption/ionization, which transfers them into the gas phase as ions.
3) Ions are then accelerated and separated by their mass-to-charge ratios to produce mass spectra, which provide information about proteins, peptides, and other molecules in biological samples.
This document discusses electrochemical sensors for detecting antibiotic residues in food. It begins with an introduction on the increasing global use of antibiotics and development of antibiotic resistance. It then discusses the working principles of electrochemical sensors and how they can be used to detect antibiotics. Specifically, it describes how electrochemical sensors use recognition elements like enzymes, antibodies, aptamers, and molecularly imprinted polymers to detect antibiotics. It also discusses using different electrode systems and materials like carbon nanotubes, nanoparticles, and graphene to improve detection. The document aims to provide an overview of developing electrochemical sensor techniques for antibiotic residue detection in food.
The document describes the development of a universal probe system using an Expanded Genetic Information System (AEGIS) that allows for simultaneous quantification and genotyping of nucleic acids via real-time PCR. RNA quantification was linear over four orders of magnitude for beta-actin mRNA and 18S rRNA. Genotyping of 176 clinical samples showed concordance between AEGIS probes and Invader® tests for factor V Leiden and prothrombin G20210A mutations in 173 samples. The AEGIS universal probe system enables rapid development of PCR assays for nucleic acid analysis.
Despite aptamers being first described in 1990, they have not been widely used in lateral flow devices. However, aptamers are receiving more attention for lateral flow applications due to performance advantages over antibodies. The top 5 reasons aptamers are better than antibodies for lateral flow assays are that they have virtually unlimited shelf-life when dried, sandwich pairs can be identified through high-throughput screening, can bind difficult targets under varied conditions, allow well-defined single-site conjugation chemistries, and can be tailored to have very fast on-rates during selection.
This document summarizes a study comparing traditional immunoassay screening followed by confirmation using solid phase extraction and gas chromatography/mass spectrometry (SPE/GC-MS) versus automated solid phase extraction high performance liquid chromatography tandem mass spectrometry (SPE/HPLC/MS/MS) for analyzing urine samples submitted for driving under the influence (DUI) cases. 106 samples previously confirmed positive by the traditional methods were re-analyzed using the automated SPE/HPLC/MS/MS system. More drugs were detected using the automated system, including 12 additional THC cases, 4 additional amphetamine cases, 6 additional benzodiazepine cases, and 3 additional cocaine cases. The automated system was able to process samples faster and at
The common labels of chemiluminescent immunoassays is alkaline phosphatase (ALP). Huateng Pharma can provides APS-5(CAS#193884-53-6) from milligram to kilogram scale.
Theoretical evaluation of shotgun proteomic analysis strategies; Peptide obse...Keiji Takamoto
This document discusses evaluating different strategies for shotgun proteomic analysis through theoretical modeling. It develops a peptide observability function based on mouse proteomic data to predict how observable peptides are by LC-MS/MS. This function is applied to theoretically digested mouse proteins using different proteases and separation techniques to evaluate their combinations and the separation profiles achieved. The results suggest SAX/trypsin and IEF/trypsin are favorable combinations that provide good separation.
This document discusses liquid chromatography-mass spectrometry (LC-MS). It begins with an introduction to LC-MS, explaining that it combines liquid chromatography separation with mass spectrometry detection. It then covers the history, principles, instrumentation, advantages, and applications of LC and MS. Key applications discussed include pharmacokinetics, proteomics, metabolomics, and drug development.
This document describes a study comparing data acquired from data-independent LC-MS to data acquired from data-dependent LC-MS/MS. The study analyzed mixtures of four proteins alone and with a complex E. coli protein digest. Each sample was run in triplicate by both acquisition methods. The data-independent LC-MS provided more comprehensive detection of precursor and product ions than the combined data-dependent LC-MS/MS experiments. Over 90% of masses detected by LC-MS/MS were also detected by data-independent LC-MS at the correct retention times with similar fragmentation patterns. The data-independent LC-MS was able to detect more components than the individual data-dependent LC-MS/MS experiments.
Bioanalytical Method Development and Validation for Simultaneous Estimation o...BRNSSPublicationHubI
The document describes the development and validation of a bioanalytical method for the simultaneous estimation of imatinib and its metabolite desmethyl imatinib in human plasma using liquid chromatography-mass spectrometry. Key steps in the method included online enrichment of the analytes followed by separation on a chromatographic column and detection using mass spectrometry. The method was validated in terms of precision, accuracy, selectivity and sensitivity. The developed method was then applied to pharmacokinetic studies of imatinib and its metabolite in patient samples.
ICP-MS detects elements instead of molecules. With the exception of a few elements (C, H, N, O and the noble gases), all elements can be detected. A specific element serves as a tag for the drug molecule of interest, thus enabling quantitation of this drug molecule in a particular matrix. The technique is highly linear and can be used quantitatively for a broad concentration range. Sample processing is relatively easy and throughput times are short, resulting in fast turnaround times.
Liquid chromatography-mass spectrometry (LC-MS) is an analytical technique that combines liquid chromatography with mass spectrometry. It has high sensitivity and selectivity. LC-MS is commonly used in pharmacokinetics studies to analyze drugs in biological samples. It is also used in proteomics to identify over 1000 proteins in complex samples like human serum. Additionally, LC-MS has various applications in drug development such as metabolite identification, quantitative bioanalysis, and quality control.
Proteomics_Chapter 3 Protein Identification.pptZaldaaZaldaa
The document provides information on learning outcomes and an overview of protein identification methods. The key learning outcomes are to understand the workflow of protein identification, identification by Western blotting and mass spectrometry, components of a mass spectrometer, and different ionization and mass spectrometry methods. It then describes in more detail the processes of protein transfer and antibody detection in Western blotting, as well as common ionization techniques like MALDI and ESI and mass analyzer types for protein identification by mass spectrometry.
Adsorptive stripping differential pulse voltammetry determination of rivastig...Pramod Kalambate
The study of graphene nanosheet (GNS)–gold nanoparticle (AuNP)–carbon paste electrode (GNS–AuNP–CPE) as
an electrochemical sensor for the determination of rivastigmine (RIV) in pharmaceuticals formulations, blood
serum, and urine samples is presented. The GNS–AuNP composite is prepared by in situ simultaneous reduction
of graphene oxide and chloroauric acid using sodiumborohydride as a reducing agent. The GNS–AuNP composite
was characterized by X-ray diffraction, UV–Vis spectroscopy, and scanning electron microscopy. Electrochemical
characterization of the GNS–AuNP–CPE electrode surfacewas carried out by cyclic voltammetry, electrochemical
impedance spectroscopy, chronocoulometry, and adsorptive stripping differential pulse voltammetry. This study
shows that oxidation of rivastigmine is facilitated at the GNS–AuNP–CPE electrode and remarkably increase in
current compared to the bare electrode due to enhanced adsorption of the former on electrode surface. Under
the optimized conditions, the peak current (Ip) is found to be proportional to the RIV concentration in the
range of 2.0 × 10–7–6.0 × 10−4 M with a detection limit of 5.3 × 10−8 M. The proposed sensor shows a very
high level of sensitivity, selectivity, and a very good reproducibility for RIV determination. A good recovery
level obtained for real samples suggests practical utility of the GNS–AuNP–CPE as an effective and reliable electrochemical
sensor for RIV detection.
The document provides an introduction to LC-MS, including:
- LC-MS combines liquid chromatography with mass spectrometry to separate and identify components in a sample.
- It can be used for qualitative analysis to determine what is in a sample and quantitative analysis to determine concentrations.
- The document reviews the basic components and workflow of LC-MS, including how the liquid chromatography separates components and the mass spectrometer ionizes and detects them.
- It provides guidance on starting up and running samples on an Agilent 6100 single quadrupole LC-MS system, including loading methods, running samples, and viewing resultant data.
Expanding Surface Plasmon Resonance Capabilities with ReichertReichertSPR
Surface Plasmon Resonance (SPR) is a widely-used label-free technique to characterize a variety of molecular interactions. SPR is an optical phenomenon that is sensitive to changes in the dielectric properties of the medium close to a metal surface. Specifically, the resonance condition is affected by changes in refractive index occurring up to 300 nm above the metal surface (Au) and thus by the material absorbed onto the metal film. Therefore, the SPR signal is a measure of the total mass concentration at the gold sensor chip surface.
Typically, a mobile molecule (analyte) is injected across an immobilized binding partner (ligand) and as the analyte binds, this mass accumulation on the sensor surface leads to an increase in refractive index, and the result is plotted as response versus time. SPR is commonly utilized by researchers to determine association/dissociation rates, affinities and thermodynamics of biomolecular interactions.
Traditionally, the interactions under study with SPR include those occurring with and between the major classes of biological macromolecules along with those involving small molecules and drugs. These classic experiments have been primarily carried with just purified samples. Reichert’s SPR systems implement a very robust fluidics arrangement that can accommodate a wide variety of sample compositions including crude samples such as lysates, whole cells and serum. In addition, Reichert’s systems are housed in an open architecture that easily allows coupling to other analytical techniques and instruments. Along with excelling at traditional biomolecular interactions, Reichert’s systems pave the way for new avenues of investigation involving crude samples and whole cells along with the ability to couple SPR to other techniques. This presentation will focus on the SPR technique and provide examples of unique applications with cells along with the possibility of interfacing SPR with other analytical methods.
http://www.reichertspr.com/webinars/general/expanding-surface-plasmon-resonance-capabilities-with-reichert/
ACCULAB Corporation is a manufacturer and supplier of laboratory and medical instruments based in New York. It incorporates 7 companies to offer a wide range of products at different price points to meet customer demands. ACCULAB aims to provide both high quality products and competitive prices with a vision of being a one-stop shop for all laboratory equipment needs through excellent customer service.
MALDI-TOF: Pricinple and Its Application in Biochemistry and BiotechnologyDevakumar Jain
The document discusses MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry. It provides a history of the development of mass spectrometry techniques. MALDI-TOF allows for the analysis of intact biomolecules like proteins and is a soft ionization method. It provides high sensitivity and mass accuracy for analyzing proteins, peptides, and other large biomolecules. The document also discusses applications of MALDI-TOF like protein identification and characterization.
This document describes a novel database search algorithm for identifying proteins from data independent acquisitions where multiple precursor ions are fragmented simultaneously. The algorithm uses an iterative process to incrementally increase selectivity, specificity, and sensitivity. It accounts for peptide retention time, ion intensities, charge states, and accurate masses of precursors and products. The algorithm was tested on simple and complex protein mixtures and validated independently, demonstrating its ability to correctly identify proteins across a wide dynamic range with high sensitivity and specificity.
Saravanan Kumar,V.Siva Reddy - Workshop on ProteomicsSARAVANAN KUMAR
1) Mass spectrometry is a technique that measures the mass-to-charge ratio of ions based on how they move in electric or magnetic fields.
2) Samples are ionized using techniques like electrospray ionization or matrix-assisted laser desorption/ionization, which transfers them into the gas phase as ions.
3) Ions are then accelerated and separated by their mass-to-charge ratios to produce mass spectra, which provide information about proteins, peptides, and other molecules in biological samples.
This document discusses electrochemical sensors for detecting antibiotic residues in food. It begins with an introduction on the increasing global use of antibiotics and development of antibiotic resistance. It then discusses the working principles of electrochemical sensors and how they can be used to detect antibiotics. Specifically, it describes how electrochemical sensors use recognition elements like enzymes, antibodies, aptamers, and molecularly imprinted polymers to detect antibiotics. It also discusses using different electrode systems and materials like carbon nanotubes, nanoparticles, and graphene to improve detection. The document aims to provide an overview of developing electrochemical sensor techniques for antibiotic residue detection in food.
The document describes the development of a universal probe system using an Expanded Genetic Information System (AEGIS) that allows for simultaneous quantification and genotyping of nucleic acids via real-time PCR. RNA quantification was linear over four orders of magnitude for beta-actin mRNA and 18S rRNA. Genotyping of 176 clinical samples showed concordance between AEGIS probes and Invader® tests for factor V Leiden and prothrombin G20210A mutations in 173 samples. The AEGIS universal probe system enables rapid development of PCR assays for nucleic acid analysis.
Despite aptamers being first described in 1990, they have not been widely used in lateral flow devices. However, aptamers are receiving more attention for lateral flow applications due to performance advantages over antibodies. The top 5 reasons aptamers are better than antibodies for lateral flow assays are that they have virtually unlimited shelf-life when dried, sandwich pairs can be identified through high-throughput screening, can bind difficult targets under varied conditions, allow well-defined single-site conjugation chemistries, and can be tailored to have very fast on-rates during selection.