This document discusses principles of immunodetection and antigen-antibody interactions. It describes how antibodies recognize antigens through epitopes and bind through various forces. Techniques like ELISA, RIA, western blotting use this interaction to detect antigens. The generation of polyclonal and monoclonal antibodies is also summarized. The document outlines clinical applications in diagnostics and therapeutics using antibodies and discusses research problems requiring immunoanalysis techniques.
Secondary Structure Prediction of proteins Vijay Hemmadi
Secondary structure prediction has been around for almost a quarter of a century. The early methods suffered from a lack of data. Predictions were performed on single sequences rather than families of homologous sequences, and there were relatively few known 3D structures from which to derive parameters. Probably the most famous early methods are those of Chou & Fasman, Garnier, Osguthorbe & Robson (GOR) and Lim. Although the authors originally claimed quite high accuracies (70-80 %), under careful examination, the methods were shown to be only between 56 and 60% accurate (see Kabsch & Sander, 1984 given below). An early problem in secondary structure prediction had been the inclusion of structures used to derive parameters in the set of structures used to assess the accuracy of the method.
Some good references on the subject:
STS stands for sequence tagged site which is short DNA sequence, generally between 100 and 500 bp in length, that is easily recognizable and occurs only once in the chromosome or genome being studied.
Ab Initio Protein Structure Prediction is a method to determine the tertiary structure of protein in the absence of experimentally solved structure of a similar/homologous protein. This method builds protein structure guided by energy function.
I had prepared this presentation for an internal project during my masters degree course.
Secondary Structure Prediction of proteins Vijay Hemmadi
Secondary structure prediction has been around for almost a quarter of a century. The early methods suffered from a lack of data. Predictions were performed on single sequences rather than families of homologous sequences, and there were relatively few known 3D structures from which to derive parameters. Probably the most famous early methods are those of Chou & Fasman, Garnier, Osguthorbe & Robson (GOR) and Lim. Although the authors originally claimed quite high accuracies (70-80 %), under careful examination, the methods were shown to be only between 56 and 60% accurate (see Kabsch & Sander, 1984 given below). An early problem in secondary structure prediction had been the inclusion of structures used to derive parameters in the set of structures used to assess the accuracy of the method.
Some good references on the subject:
STS stands for sequence tagged site which is short DNA sequence, generally between 100 and 500 bp in length, that is easily recognizable and occurs only once in the chromosome or genome being studied.
Ab Initio Protein Structure Prediction is a method to determine the tertiary structure of protein in the absence of experimentally solved structure of a similar/homologous protein. This method builds protein structure guided by energy function.
I had prepared this presentation for an internal project during my masters degree course.
Current trends in pseduogene detection and characterizationShreya Feliz
This presentation gives the insight of the current trends in detecting and characterizing Pseudogenes. Pseudogenes detection by bioinformatics may enhance the understanding of Pseudogenes and take research to the next step.
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
The S1 nuclease was extracted from Aspergill suoryzae. The S1 nuclease is a specific
single-stranded endonuclease. It can degrade single-stranded DNA and
single-stranded RNA to produce 5'-single-stranded nucleotides or oligonucleotides.
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Current trends in pseduogene detection and characterizationShreya Feliz
This presentation gives the insight of the current trends in detecting and characterizing Pseudogenes. Pseudogenes detection by bioinformatics may enhance the understanding of Pseudogenes and take research to the next step.
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
The S1 nuclease was extracted from Aspergill suoryzae. The S1 nuclease is a specific
single-stranded endonuclease. It can degrade single-stranded DNA and
single-stranded RNA to produce 5'-single-stranded nucleotides or oligonucleotides.
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Get expert antibody phage display and antibody library services from professional dealers. Creative Biolabs is right place that focuses on phage display service using the advanced technology.
Presentation of Frank Hills in 1st International Antibody Validation Forum 2014St John's Laboratory Ltd
After graduating with an honours degree in Biochemistry Dr Hills worked for several years as a Clinical Scientist at St Bartholomew's hospital in London. He was awarded his PhD in 2002 from the faculty of Medicine at Queen Mary University of London. He continued his interest in reproductive science at Imperial College London where he worked as a postdoctoral researcher before joining Middlesex in 2004 as a lecturer. Dr Hills has published many high profile original research articles on various aspects of obstetric pathology including pre-eclampsia, recurrent miscarriage, preterm labour and fetal distress as well as several articles in the area of assisted reproduction. Currently, he is research interests include investigating the role of glycosaminoglycans and proteoglycans on the development of placental pathology and breast cancer. Dr Hills teaches a range of topics in biomedical science including clinical biochemistry, cellular and developmental biology as well as statistical analysis. He is author of around 30 peer-reviewed scientific articles and has refereed manuscripts for a variety of journals in the area of reproduction and endocrinology.
For more details about 1st international antibody validation forum please check on http://www.stjohnslabs.com/ac_cms/blog
Exploring the First Line of Defense - Research Tools for Innate Immnity: Host...QIAGEN
The innate immune system executes crucial and unique functions for host defense against infection. This slidedeck provides an overview of the most important cellular and molecular components of innate immunity and discusses their functions in a variety of disease states. Research technologies are also introduced for exploring innate immune activity in your system through profiling of gene expression, cytokine production and signal transduction pathway analysis, all in the context of current literature.
Learn about novel cell-based assays that enable improved immunotherapy drug development. See case studies utilizing checkpoint receptors such as PD-1, VISTA, and NIK.
Climate Impact of Software Testing at Nordic Testing DaysKari Kakkonen
My slides at Nordic Testing Days 6.6.2024
Climate impact / sustainability of software testing discussed on the talk. ICT and testing must carry their part of global responsibility to help with the climat warming. We can minimize the carbon footprint but we can also have a carbon handprint, a positive impact on the climate. Quality characteristics can be added with sustainability, and then measured continuously. Test environments can be used less, and in smaller scale and on demand. Test techniques can be used in optimizing or minimizing number of tests. Test automation can be used to speed up testing.
Elevating Tactical DDD Patterns Through Object CalisthenicsDorra BARTAGUIZ
After immersing yourself in the blue book and its red counterpart, attending DDD-focused conferences, and applying tactical patterns, you're left with a crucial question: How do I ensure my design is effective? Tactical patterns within Domain-Driven Design (DDD) serve as guiding principles for creating clear and manageable domain models. However, achieving success with these patterns requires additional guidance. Interestingly, we've observed that a set of constraints initially designed for training purposes remarkably aligns with effective pattern implementation, offering a more ‘mechanical’ approach. Let's explore together how Object Calisthenics can elevate the design of your tactical DDD patterns, offering concrete help for those venturing into DDD for the first time!
The Metaverse and AI: how can decision-makers harness the Metaverse for their...Jen Stirrup
The Metaverse is popularized in science fiction, and now it is becoming closer to being a part of our daily lives through the use of social media and shopping companies. How can businesses survive in a world where Artificial Intelligence is becoming the present as well as the future of technology, and how does the Metaverse fit into business strategy when futurist ideas are developing into reality at accelerated rates? How do we do this when our data isn't up to scratch? How can we move towards success with our data so we are set up for the Metaverse when it arrives?
How can you help your company evolve, adapt, and succeed using Artificial Intelligence and the Metaverse to stay ahead of the competition? What are the potential issues, complications, and benefits that these technologies could bring to us and our organizations? In this session, Jen Stirrup will explain how to start thinking about these technologies as an organisation.
DevOps and Testing slides at DASA ConnectKari Kakkonen
My and Rik Marselis slides at 30.5.2024 DASA Connect conference. We discuss about what is testing, then what is agile testing and finally what is Testing in DevOps. Finally we had lovely workshop with the participants trying to find out different ways to think about quality and testing in different parts of the DevOps infinity loop.
Le nuove frontiere dell'AI nell'RPA con UiPath Autopilot™UiPathCommunity
In questo evento online gratuito, organizzato dalla Community Italiana di UiPath, potrai esplorare le nuove funzionalità di Autopilot, il tool che integra l'Intelligenza Artificiale nei processi di sviluppo e utilizzo delle Automazioni.
📕 Vedremo insieme alcuni esempi dell'utilizzo di Autopilot in diversi tool della Suite UiPath:
Autopilot per Studio Web
Autopilot per Studio
Autopilot per Apps
Clipboard AI
GenAI applicata alla Document Understanding
👨🏫👨💻 Speakers:
Stefano Negro, UiPath MVPx3, RPA Tech Lead @ BSP Consultant
Flavio Martinelli, UiPath MVP 2023, Technical Account Manager @UiPath
Andrei Tasca, RPA Solutions Team Lead @NTT Data
Enhancing Performance with Globus and the Science DMZGlobus
ESnet has led the way in helping national facilities—and many other institutions in the research community—configure Science DMZs and troubleshoot network issues to maximize data transfer performance. In this talk we will present a summary of approaches and tips for getting the most out of your network infrastructure using Globus Connect Server.
Alt. GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using ...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
State of ICS and IoT Cyber Threat Landscape Report 2024 previewPrayukth K V
The IoT and OT threat landscape report has been prepared by the Threat Research Team at Sectrio using data from Sectrio, cyber threat intelligence farming facilities spread across over 85 cities around the world. In addition, Sectrio also runs AI-based advanced threat and payload engagement facilities that serve as sinks to attract and engage sophisticated threat actors, and newer malware including new variants and latent threats that are at an earlier stage of development.
The latest edition of the OT/ICS and IoT security Threat Landscape Report 2024 also covers:
State of global ICS asset and network exposure
Sectoral targets and attacks as well as the cost of ransom
Global APT activity, AI usage, actor and tactic profiles, and implications
Rise in volumes of AI-powered cyberattacks
Major cyber events in 2024
Malware and malicious payload trends
Cyberattack types and targets
Vulnerability exploit attempts on CVEs
Attacks on counties – USA
Expansion of bot farms – how, where, and why
In-depth analysis of the cyber threat landscape across North America, South America, Europe, APAC, and the Middle East
Why are attacks on smart factories rising?
Cyber risk predictions
Axis of attacks – Europe
Systemic attacks in the Middle East
Download the full report from here:
https://sectrio.com/resources/ot-threat-landscape-reports/sectrio-releases-ot-ics-and-iot-security-threat-landscape-report-2024/
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Removing Uninteresting Bytes in Software FuzzingAftab Hussain
Imagine a world where software fuzzing, the process of mutating bytes in test seeds to uncover hidden and erroneous program behaviors, becomes faster and more effective. A lot depends on the initial seeds, which can significantly dictate the trajectory of a fuzzing campaign, particularly in terms of how long it takes to uncover interesting behaviour in your code. We introduce DIAR, a technique designed to speedup fuzzing campaigns by pinpointing and eliminating those uninteresting bytes in the seeds. Picture this: instead of wasting valuable resources on meaningless mutations in large, bloated seeds, DIAR removes the unnecessary bytes, streamlining the entire process.
In this work, we equipped AFL, a popular fuzzer, with DIAR and examined two critical Linux libraries -- Libxml's xmllint, a tool for parsing xml documents, and Binutil's readelf, an essential debugging and security analysis command-line tool used to display detailed information about ELF (Executable and Linkable Format). Our preliminary results show that AFL+DIAR does not only discover new paths more quickly but also achieves higher coverage overall. This work thus showcases how starting with lean and optimized seeds can lead to faster, more comprehensive fuzzing campaigns -- and DIAR helps you find such seeds.
- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
Why You Should Replace Windows 11 with Nitrux Linux 3.5.0 for enhanced perfor...SOFTTECHHUB
The choice of an operating system plays a pivotal role in shaping our computing experience. For decades, Microsoft's Windows has dominated the market, offering a familiar and widely adopted platform for personal and professional use. However, as technological advancements continue to push the boundaries of innovation, alternative operating systems have emerged, challenging the status quo and offering users a fresh perspective on computing.
One such alternative that has garnered significant attention and acclaim is Nitrux Linux 3.5.0, a sleek, powerful, and user-friendly Linux distribution that promises to redefine the way we interact with our devices. With its focus on performance, security, and customization, Nitrux Linux presents a compelling case for those seeking to break free from the constraints of proprietary software and embrace the freedom and flexibility of open-source computing.
2. Aims and Objectives
• Basis of antigen-antibody interaction
• Conceptualise the different techniques
based on this interaction
• Examples of clinical application
• Research problems requiring
immunoanalyses
3. Role of antibodies
• Protect against
– Viral infections
– Bacterial infections
– Foreign bodies
• Antigens
• Deleterious in
– Autoimmune diseases
• Reumathoid arthritis Lupus
• Type 1 diabetes Croh’n disease
– Graft rejection
4. Antigen-antibody interaction
• Antigen: foreign molecules that generate antibodies or any
substance that can be bound specifically by an antibody
molecule
– Proteins, sugars, lipids or nucleic acids
– Natural or synthetic
• Antibody: molecules (protein) responsible for specific
recognition and elimination (neutralization) of antigens
– Different structures (7-8 classes in mammals)
– Powefull research tools for basic research, clinical applications and
drug design
5. Antigenic determinants
• An antibody will recognize
– Epitope: defined segment of an antigen
– Immunoreactivity of epitopes may depend on primary,
secondary, tertiary or quaternary structure of an antigen
– Variability of epitopes depends on the species
• Antibodies are antigen themselves
6. Nature of binding forces
• Hydrogen bonding
– Results from the formation of hydrogen bridges between appropriate atoms
• Electrostatic forces
– Are due to the attraction of oppositely charged groups located on two protein side
chains
• Van der Waals bonds
– Are generated by the interaction between electron clouds (oscillating dipoles)
• Hydrophobic bonds
– Rely upon the association of non-polar, hydrophobic groups so that contact with water
molecules is minimized (may contribute up to half the total strength of the antigen-antibody
bond)
9. Antigen-antibody affinity
The affinity with which antibody binds antigen results from a balance
between the attractive and repulsive forces. A high affinity antibody implies
a good fit and conversely, a low affinity antibody implies a poor fit and a
lower affinity constant
13. Generation of antibodies:
polyclonal vs monoclonal
• Host animals ca be used to raise antibodies
against a given antigen
• Slected clones from a polyclonal each recognizing
a single epitope can be fused to a tumor cell
(hybridoma) to proliferate indefinitely
14. Laboratory use of antibodies
• Quantitation of an antigen
– RIA, Elisa
• Identification and characterization of protein antigens
– Immunoprecipitation
– Western blotting
• Cell surface labelling and separation
• Localisation of antigens within tissues or cells
• Expression librairies
• Phage display
16. • General equation for a
dose response curve
• It shows response as a
function of the logarithm
of concentration
• X is the logarithm of
agonist concentration
and Y is the response
• Log EC50 is the
logarithm of the EC50
(effective concentration,
50%)
• IC50 (inhibitory conc.)
Sigmoidal dose response curve
10%
90%
30. Clinical use of antibodies
• Diagnostic
– Detection of peptides and other molecules in various diseases
• Endocrine diseases: hyperinsulinemia, diabetes, hyperparatyroidism
• Tumor antigens (p53 tumor suppressor, PSA, a-foetoprotein)
• Antibodies against viral proteins (AIDS, hepatitis)
• Therapeutic
– Neutralizing antibodies
• Anti-Erbb2 for breast and ovarian cancer
• Anti-CD20 for B-cell non-Hodgkin's lymphoma
• Experimental
– Drug screening (phage display)
31. Detection of HIV proteins by WB
gp160 viral envelope precursor (env)
gp120 viral envelope protein (env) binds to CD4
p31 Reverse Transcriptase (pol)
p24 viral core protein (gag)
32. • Phosphorylation and dephosphorylation affect
the structure and activity of proteins
• Cellular signalling is characterized by cascades
of phosphorylation
• Kinases and phosphatases maintain
phosphorylated/dephosphorylated state of
proteins
• Phospho/Tyrosine/Threonine/ Serine
Phosphospecific antibodies to study
cellular signaling
37. Localization of BFP- and RFP-C/EBP protein expressed in mouse 3T3 cells using
2p-FRET microscopy. The doubly expressed cells (BFP-RFP-C/EBP) were excited
by 740 nm and the donor (A) and acceptor (B) images of proteins localized in the
nucleus of a single living cell were acquired by single scan
Localization of CEBP by FRET
41. Phage display: Ab production
Originally developped to produce monoclonal
antibodies, phage display is a simple yet
powerful technology that is used to rapidly
characterize protein-protein interactions from
amongst billions of candidates. This widely
practiced technique is used to map antibody
epitopes, create vaccines and to engineer
peptides, antibodies and other proteins as both
diagnostic tools and as human therapeutics
42. Clinical applications
• Neutralizing antibodies
– Antidotes and antivenin (snake & spider bites)
– Tumor antigens ErbB-2, melanoma and T-cell leukemia,
antibodies coupled to toxins
– Autoimmune antibodies, cytokines TNF-a
– Antisera aigainst virus, bateria and toxins (vaccine)
– Anti IgE and IgM for allegies (experimental)
– Quantitation of blood peptides (hormones metabolites)
• Activating antibodies
– Complement activating for uncontrolled bleeding (hemophilia)
44. • Identification of signaling pathways
– Protein modifications
– Signaling partners
• Activity of drugs (lead compounds)
• Lack of specific molecules
– Specific ligands (side effects)
– New antibodies
Research problems requiring
immunoanalyses
45. The problems of chemotherapy
Chemotherapy/
radiotherapy
Sensors
Transducers
Cytoplasmic/Nuclear effectors
Chromatin
Structure
Transcription
DNA repair
Cell cycle
checkpoints
Apoptosis
Drug resistance arising
from sensor/transducer
defects
Drug resistance arising
from effector defects
DNA Damage
Drug resistance arising
from altered drug
delivery to target