MALDI-TOF mass spectrometry is a technique used to analyze proteins. It works by ionizing protein samples using a laser and then measuring the time it takes for the ions to travel through a flight tube, which allows calculating the mass-to-charge ratio. The sample is mixed with an absorbing matrix and dried on a target plate before being ionized by a laser pulse. Ions are accelerated through a flight tube and reach a detector, with lighter ions traveling faster and reaching it first. The time of flight is converted to a mass spectrum, allowing identification of proteins in the sample. MALDI-TOF provides sensitive, high-throughput protein analysis and is widely used in fields like proteomics, microbiology,
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
mass spectrometry, also called mass spectroscopy, analytic technique by which chemical substances are identified by the sorting of gaseous ions in electric and magnetic fields according to their mass-to-charge ratios.
What is ICH- GCP?
Why is GCP important?
Outline the goals of GCP
Provide a historical perspective on GCP
WHO Principles of GCP
Principles: Defines, Application & Implementation.
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This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
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Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
A Strategic Approach: GenAI in EducationPeter Windle
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This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
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Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
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June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
2. INTRODUCTION
• Proteome – complete set of proteins encoded in a
genome
• Proteomics
• Protein identification requires their separation by
methods :_
1. IEF
2. SDS- gel electrophoresis
3. Mass spectroscopy (MS)
3. MASS SPECTROSCOPY
• Separation of charged molecules/ molecules
according to their mass to charge
• Determines relative molecular mass (Mr)
• High resolution, precision, & sensitivity
• Requires charged gaseous molecules for its analysis
4. COMPONENTS OF MASS
SPECTROSCOPY
• Ionization source
1. Matrix assisted laser desorption/ ionization – time of
flight(MALDI)
2. Electrospray (ES)
• Analyser m/z : TOF, Magnet,..
• Detector : Photomultiplier, Electron multiplier, ..
5. COMPONENTS OF MASS
SPECTROSCOPY
Inlet Detector
Data
System
Mass
Analyzer
High Vacuum System
Ion
Source
Time of flight
(TOF)
Quadraupole
Ion Trap
Magnetic Sector
FTMS
Turbo
molecular
pumps
HPLC
Flow
injection
Sample plate
Microchannel
Plate
Electron
Multiplier
Hybrid with
photomultiplier
PC
Sun SPARK
Station
DEC Station
6. MALDI - TOF
• Synonyms : MALDI, TOF – MS
• MALDI- Term was coined in 1985 by Franz Hillenkamp,
Michael Karas
• Principle : velocity of ions depend on its mass & energy,
the time taken by an ion to travel a specified distance
(TOF)
• Use : MALDI ionization & TOF of the ions to reach the
detector as a parameter to measure m/z ratio.
• Sample excitation – from the energy of a laser transferred
via a UV light absorbing matrix
7. MATRIX
• Conjugated organic compound or weak organic acid
• Weak organic acid – derivative of cinnamic acid &
dihydroxybenzoic acid
• Eg : sinapinic acid- proteins (10Da)
• Usually mixed with sample
• Absorbs maximum light at a wavelength (λ) of the laser,
typically a N2 laser of 337nm or yytrium – aluminium
garnet (Nd-YAG) AT 335nm.
• Acts as an absorbing media for the UV light
8. WORKING
Sample
(1 – 10 pmolmm-3)
Mixing Excess matrix
Sample – matrix
Dried on target plate
Co- crystallization
UV rays
Desorption
Pulses of laser light
Rapid excitation
9. Rapid heating of the region
Matrix & analyte ions ejection
Into
Gas phase
Results in
Explotion of sample region into high vacuum
Gas phase protonated molecules
Enter
TOF
12. SAMPLE CONCENTRATION FOR MATRIX
• Maximum sensitivity if samples a diluted to a specific
concentration range
• For unknown sample concentration a dilution series may be
needed
• Sample loaded as a spot
• Peptides & proteins give best spectra (10pmolmm-3)
• Especially glycoproteins (10pmolmm-3)
• Oligonucleotides- better spectra at 10 - 100pmolmm-3
• Polymers - 100pmolmm-3
13. MALDI ADVANTAGES
• Gentle Ionization technique
• High molecular weight analyte can be ionized
• Molecule need not be volatile
• Sub-picomole sensitivity easy to obtain
• Wide array of matrices
14. TOF
• Best type of mass analyser to couple to MALDI
• Unlimited mass range
• Macromolecules of Mr > 400,000 – accurately measured
• 100 pulses of laser light (10nanoseconds)
• Result – seen as a good spectrum
• Camera – tracks the laser beam around the MALDI
15. TOF ADVANTAGES
• All ions detected at once
• High mass accuracy and resolving power possible
• Reasonable performance for cost
▫ <5 ppm mass accuracy and >20,000 resolving power
commercially available
• High mass, low charge ions not a problem
▫ Theoretically unlimited mass range
18. LINEAR TOF MS
• Uses MALDI & TOF
• Samples are deposited on a metal substrate (100)
Analyte spots
laser
Short burst of ions
Ion acceleration to a fixed KE
Ions travel down a flight tube
19. DE MALDI – TOF MS
• Delayed extraction TOF MS
• Mass dependent
• Used for DNA sequencing
• Lacks resolution & sentivity to analyze complex
mixtures
20. RE TOF MS
• Reflectron TOF MS
• Single stage or dual stage reflectron – at the end of
flight tube
• High resolution
• Reflectron – compensate for the difference in the
TOF of ions with same m/z ratio, but same KE
21. DELAYED EXTRACTION(DE)
• Extraction of ions generated by a high electrostatic field
occurs
• Ions from different spots have different energy
• Energy spreads, so broadens the peak corresponding to each
ion (low accuracy)
• If extraction is delayed until all ions are formed, spread
minimizes (high accuracy) : DE
• Here extraction occurs by high voltage.
• Lengthening time of delay controls ion fragmentation degree.
22.
23. POST SOURCE DECAY
• Ion extraction results in ion fragmentation
• Biological molecules gives rise to molecules that
have been extracted before this dissociation is
complete
• Fragment ion’s velocity = precursor velocity
• Use of reflector overcomes this.
• Give structural information
24. MALDI – TOF INSTRUMENT
COMPONENTS
• Sample target plate
• High vacuum chamber
• Camera
• Laser beam source
• Clock
• Flight tube
• Deflector
• Data system
• Reflectron
26. WORKING OF THE COMPONENTS
Sample-matrix
Target plate
High vacuum chamber
Camera tracks laser beam
Irradiation with laser pulses
27. Clock (on)
Measures TOF
EF
Acceleration of ions to same kinetic energy
Ions fly through the flight tube
Ion separation (mass)
Ions strike detector
Data system
(controls instrument parameters, acquires signal v/s time &
process the data )
28. TYPES OF MALDI SAMPLE PLATES
1. 100 well stainless steel flat plates
2. Four – hundred – spot Teflon – coated plates
3. Gold - coated plates
29. DETECTORS
• Ions from MS impinge on its surface
• Has a neutral charge on surface
• I flows through its surface
• I gets amplified
• I gets converted to signal
• Signal processed by a computer
• TIC – I 1 + I2 + ……..In
• TIC – to measure during online MS
30. REFLECTRON
• Reflector
• An ion mirror that provides higher resolution
• Increases overall path length for an ion
• Corrects minor variation in the energy spread of ions
of the same mass.
• Has a gradient electric field
• The depth to which ions will penetrate this field,
before reversal of direction of travel, depends upon
their energy.
31. • Higher energy ions travels more & vice versa.
• Thus, TOF gets focussed
• Neutral fragments - unaffected by deflection
• Improves resolution & mass accuracy
• Allows structure & sequence information collection by
PSD analysis.
• Focuses charged fragments of a specific range of m/z
• So, a number of spectra are run at different settings &
stiched together to generate composite spectrum.
