This article is aimed at a brief introduction to phage display technology for production of single-domain Abs (dAbs), popularly also called ‘nanobodies’, and then to discuss their diagnostic applications.
DNA Nanotechnology: Concept and its Applications
DNA Nanotechnology # Various 2 and 3 dimensional shapes of DNA nanotechnology # DNA Origami # with their application and Future scope
DNA Nanotechnology: Concept and its Applications
DNA Nanotechnology # Various 2 and 3 dimensional shapes of DNA nanotechnology # DNA Origami # with their application and Future scope
pBluescript is an example of a combination between plasmids and phages (phagemids).
Phagemids represent a hybrid type of class of vectors that serve to produce single-stranded DNA.
Metagenomics is the study of metagenome, genetics material, recovered directly from environmental sample such as soil, water or faeces.
Metagenomics is based on the genomics analysis of microbial DNA directly
from the communities present in samples
Metagenomics technology – genomics on a large scale will probably lead to great advances in medicine, agriculture, energy production and bioremediation.
Metagenomics can unlock the massive uncultured microbial diversity present in the environment for new molecule for therapeutic and biotechnological application.
Metagenomic studies have identified many novel microbial genes coding for metabolic pathways such as energy acquisition, carbon and nitrogen metabolism in natural environments that were previously considered to lack such metabolism
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Hilde Revets, Senior Research Fellow, Ablynx
Presentation at EIPG – VAPI-UPIP Symposium “Biotech and Advanced Therapies: Challenges and Opportunities” at the Faculty of Medicine and Pharmacy, Campus Jette, Vrije Universiteit van Brussel, Brussels 2013.
pBluescript is an example of a combination between plasmids and phages (phagemids).
Phagemids represent a hybrid type of class of vectors that serve to produce single-stranded DNA.
Metagenomics is the study of metagenome, genetics material, recovered directly from environmental sample such as soil, water or faeces.
Metagenomics is based on the genomics analysis of microbial DNA directly
from the communities present in samples
Metagenomics technology – genomics on a large scale will probably lead to great advances in medicine, agriculture, energy production and bioremediation.
Metagenomics can unlock the massive uncultured microbial diversity present in the environment for new molecule for therapeutic and biotechnological application.
Metagenomic studies have identified many novel microbial genes coding for metabolic pathways such as energy acquisition, carbon and nitrogen metabolism in natural environments that were previously considered to lack such metabolism
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Hilde Revets, Senior Research Fellow, Ablynx
Presentation at EIPG – VAPI-UPIP Symposium “Biotech and Advanced Therapies: Challenges and Opportunities” at the Faculty of Medicine and Pharmacy, Campus Jette, Vrije Universiteit van Brussel, Brussels 2013.
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.
Mycoplasmas are the smallest and simplest self-replicating bacteria .These microorganisms lack a rigid cell wall and are bound by a single membrane, the plasma membrane. The lack of a cell wall is used to distinguish these microorganisms from ordinary bacteria and to include them in a separate class named Mollicutes. Most human and animal mollicutes are Mycoplasma and Ureaplasma species of the family Mycoplasmataceae. Because mycoplasmas have an extremely small genome, these organisms have limited metabolic options for replication and survival. The smallest genome of a self-replicating organism known at present is the genome of Mycoplasma genitalium (0.58 Mb). Owing to their limited biosynthetic capabilities, most mycoplasmas are parasites exhibiting strict host and tissue specificities. The mycoplasmas enter an appropriate host in which they multiply and survive for long periods of time. These microorganisms have evolved molecular mechanisms needed to deal with the host immune response and the transfer and colonization in a new host. These mechanisms include mimicry of host antigens, survival within phagocytic and nonphagocytic cells, and generation of phenotypic plasticity. The major question is whether mycoplasmas cause damage to the host cells and to what extent the damage is clinically apparent. Mycoplasmas have long resisted detailed analyses because of complex nutritional requirements, poor growth yields, and a paucity of useful genetic tools. Although questions still far outnumber answers, significant progress has been made in identifying the mechanisms by which mycoplasmas interact and damage eukaryotic host cells. Many animal mycoplasmas depend on adhesion to host tissues for colonization and infection. In these mycoplasmas adherence is the major virulence factor, and adherence-deficient mutants are avirulent. Primary interactions between the host and mycoplasma cells occur through cell surface adhesins produced by the mycoplasma. The adhesins have been characterized in only a limited number of mycoplasmas, and while there are homologs of the characterized adhesins in some other mycoplasmas, it is clear that different molecules and structures may be involved in adhesion in different species.
Antibody screening is more helpful in identifying the
highly productive cells and is more significant in the
form of laboratory techniques. Complete solution on screening
of antibodies can be effectively offered only
by such kinds of reputed medical institutions.
For more details please visit
http://www.axiomxinc.com/
Presentation from the 2014 Waterloo iGEM team at the Giant Jamboree in Boston. Read more about Staphylocide, our microbe engineered to silence antiobiotic resistance, on our 2014 wiki: http://2014.igem.org/Team:Waterloo.
This presentation is also available on the iGEM website: http://2014.igem.org/files/presentation/Waterloo_Championship.pdf
This master's seminar presentation speaks about the role of bacteriophage in the management of different plant diseases.
It deals with the history and discovery of bacteriophages up to current research studies and usage.
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses.
Monoclonal antibodies are important reagents used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer.
These antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the subject of study.
The Karolinska Institute (KI) is the largest centre for medical education and research in Sweden and the home of the Nobel Prize in Physiology or Medicine.
KI consists of 22 departments and 600 research groups dedicated to improving human health through research and higher education.
The role of the Kohonen/Grafström team has been to guide the application, analysis, interpretation and storage of so called “omics” technology-derived data within the service-oriented subproject “ToxBank”.
History
Host pathogen interaction
R gene
Molecular techniques for detection of plant pathogens
Role of molecular techniques in resistance breeding Deployment of R genes and linked markers
Transgenic approaches in plant protection
Conclusion
Evaluating Ozoralizumab (ATN-203) as a Novel Biotherapeutic Agent for the Tre...jake9606
Poster Presentation which focuses on the background, production and direction of novel Nanobody Technology as a potential biotherapeutic. Technology has been developed by Ablynx (Belgium) and has many benefits particularly in production compared to Antibodies.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
3. Conventional antibody and their
engineering
CH2
CH3
Camel
Paratope
VHH
Hinge
DOMAIN ANTIBODY
(NANOBODY)
(Ag binding site)
Fig. 3 Camelid heavy chain antibody (HCAb) nanobody/single domain
Abs & their derivatives that can be produced by rDNA techniques
. and
CHOs
CAMELID HCAb
Human framework regions
HUMANIZED NANOBODY
BISPECIFIC NANOBODY BIFUNCTIONAL NANOBODY
Ag1
binding
paratope
Ag2binding
paratope
Nanobody “magic bullet”
Enzyme/Radionuclide/toxin
conjugate
Paratope
4. Heavy chain antibodies (hcAbs)
Camel sera also contain
antibodies devoid of light chains
and CH1
(Hamers-Casterman et al. 1993)
90 KDa
Also in Camelidae family (e.g.,
llamas and alpacas)
Immunoglobulin new antigen
receptor, (Ig-NAR) discovered in
cartilaginous fish
(Greenberg et al.1995)
5. Nanobodies (Nb)
Single domain antibodies (SdAb)
The recombinant antigen-specific, single-domain of the Heavy
chain of the Heavy-chain antibody (VHH) with dimensions in the
nanometer range.
6. VHH domain of a camelid heavy chain
antibody
6
Muyldermans, 2013
16. Affinity maturation
• Process to improve antibody affinity for an
antigen.
• In vivo, by somatic hypermutation and clonal
selection.
• In vitro, in the laboratory affinity maturation
can be obtained by mutation and selection.
17. Expression
17
Bacteria- Periplasmic Nb production 1 to 10 mg/l
Fungi - from <1 to >100 mg/l
Mammalian cell lines - when eukaryotic protein folding
machinery is required
Plants - free of possible contamination with human
and animal pathogens
Nb-Fc antibodies in soyabean as feed for oral
passive immunisation De Meyer et al.., 2014
18. Immunogenicity
• Nanobodies tested to date
do not appear to show any
unexpected levels of
immunogenicity
• VHH domains from which
Nanobodies are derived
show a high degree of
homology with human VH
domains
21. Pathogen and toxin detection
• Distinguish between Brucella and Yersinia infections in livestock,
mAbs have failed. (Abbady, et al. 2012)
• Anti-caffeine Nbs for the quantification of caffeine in hot
beverages. (Ladenson et al. 2006)
• Nanobodies against three toxins, staphylococcal enterotoxin B and
botulinum toxin A complex toxoid, have been isolated by panning a
semi-synthetic shark dAb display library. (Liu et al., 2007).
• Sandwich assays using these dAbs as the reporter antibody were
developed to demonstrate their utility for future sensor applications
(Liu et al., 2007).
23. Molecular imaging
23
Chakravarty et al., 2014
Fast extravasation, good
tumour penetration, and
rapid renal clearance of
excess tracer
Radiolabelled Nbs
24. Nanobodies as Therapeutics
24
Pathogen targeting
Bacteria/Phages
Viruses
Fungi/ Parasites
Crossing blood brain barrier
Inhibition of enzymes, cell surface and other soluble
proteins
Cancer therapeutics
Auto immune diseases
Others
25. Targeting bacteria and phages
25
Transformed lactobacilli with VHH against Streptococcus
mutans, in dental caries.
Reducing the bacterial resistance to antibiotics by VHHs with
beta-lactamase inhibitory effect.
Preventing the infection of Lactococcus lactis cultures by the p2
bacteriophage by Nbs against a phage tail protein Engineering
(an interesting approach for vaccinating microorganisms of
biotechno-logical interest)
(Wesolowski et al. 2009)
26. Targeting fungi, protozoans, toxins
26
Against a cell wall protein of Malassezia furfur, a fungus
implicated in dandruff –used in shampoo formulation
Trypanosomes (oligomannose cryptic epitope)
Scorpion, snake venom
(Baral et al. 2006)
27. Crossing Blood Brain Barrier
27
Nanobody FC5, binding to a putativ
a(2,3)-sialoglycoprotein receptor
Apolipoprotein E (ApoE) binds to
low density lipoprotein receptor-
related protein 1 (LRP1) inducing
transcytosis
Transferrin receptor (TrfR)
Shifting the isoelectric point (pI) of
(Rissiek et al. 2014)
28. Cancer therapeutics
28
platform A : naked nanobodies
platform B: nanobodies fused
to effector domains
platform C : nanobodies
decorating the surface of
nanoparticles
Kijanka et al 2015
29. Nanobodies as Research Tools
29
Protein purification and immunoprecipitation
crystallization and structural determination of
challenging targets
30. Protein purification and
immunoprecipitation
30
As protein purification ligands
Stability ensures a high column regeneration capacity
Nbs with an anti-human IgG specificity for depletion of
IgGs from blood (BAC BV/Life Technologies)
A new protein affinity tag of only four amino acids (EPEA)
Nguyen-Duc et al.., 2013
31. Crystallization chaperones
• Determining protein structures by X-ray crystallography is
difficult for ‘high hanging fruits’ such as membrane
proteins and large protein complexes.
• Nanobodies facilitated crystallization of many proteins
32. SUMMARY
32
Single domain antibodies from heavy chain antibodies
An accessible and streamlined protocol available
Recombinant Nbs are well expressed, highly robust, and easy to
engineer
Prefer to interact with cavities on the surface of their antigen
Applications in research, diagnostic and therapeutics