The heart of the fermentation or bioprocess technology is the Fermentor or Bioreactor. A bioreactor is basically a device in which the organisms are cultivated to form the desired products. it is a containment system designed to give right environment for optimal growth and metabolic activity of the organism.
A fermentor usually refers to the containment system for the cultivation of prokaryotic cells, while a bioreactor grows the eukaryotic cells (mammalian, insect cells, etc).
This PPT dicusses about the Stirred Tank Bioreactor and its features mainly used in Fermentation process.
Useful for students doing their Bachelor's in Life Science
The heart of the fermentation or bioprocess technology is the Fermentor or Bioreactor. A bioreactor is basically a device in which the organisms are cultivated to form the desired products. it is a containment system designed to give right environment for optimal growth and metabolic activity of the organism.
A fermentor usually refers to the containment system for the cultivation of prokaryotic cells, while a bioreactor grows the eukaryotic cells (mammalian, insect cells, etc).
This PPT dicusses about the Stirred Tank Bioreactor and its features mainly used in Fermentation process.
Useful for students doing their Bachelor's in Life Science
molecular biology phage vector, full lifecycle and all necessary information regarding lambda phage, it contain 2 types that is insertion and replacement.
What is PCR ? What is Real Time PCR ? Polymerase Chain Reaction ? What is Reverse Transcriptase Enzyme ?
Presented By:
Bharat Bhushan Negi
M.Tech. Biotechnology
IIT Guwahati
This presentation covers the introduction to Insect Cell Culture. Also covers its general information about cell culture practices followed in the lab. It covers culture media, the source of cells for culture and examples of the cell line with their culture conditions.
A guide to lateral flow immunoassay developmentExpedeon
This latest presentation on lateral flow immunoassay development will provide a general overview of lateral flow assays, take you through the components of a typical lateral flow test strip, and will provide you with detail on the different detection methods which are employed. We will also describe how our products and custom services can greatly simplify the development of your lateral flow assay.
To find out more about Innova Biosciences' lateral flow assay development services visit our website:
https://www.innovabiosciences.com/b2b/lateral-flow-assay-development-service.html
molecular biology phage vector, full lifecycle and all necessary information regarding lambda phage, it contain 2 types that is insertion and replacement.
What is PCR ? What is Real Time PCR ? Polymerase Chain Reaction ? What is Reverse Transcriptase Enzyme ?
Presented By:
Bharat Bhushan Negi
M.Tech. Biotechnology
IIT Guwahati
This presentation covers the introduction to Insect Cell Culture. Also covers its general information about cell culture practices followed in the lab. It covers culture media, the source of cells for culture and examples of the cell line with their culture conditions.
A guide to lateral flow immunoassay developmentExpedeon
This latest presentation on lateral flow immunoassay development will provide a general overview of lateral flow assays, take you through the components of a typical lateral flow test strip, and will provide you with detail on the different detection methods which are employed. We will also describe how our products and custom services can greatly simplify the development of your lateral flow assay.
To find out more about Innova Biosciences' lateral flow assay development services visit our website:
https://www.innovabiosciences.com/b2b/lateral-flow-assay-development-service.html
Semi Automated Low-throughput Workflow for Microbial Analyses of Human StoolQIAGEN
The gut microbiota composition changes dramatically throughout aging and disease. A healthy gut microbiota is typically characterized by large bacterial taxonomic diversity and functional capacity, whereas frailty and aging are associated with loss of diversity and expansion of more pathogenic bacterial species. However, in order to accurately profile changes in microbial communities, the reproducible isolation of high-quality DNA is an important step. Automation enables reliable and reproducible isolation of DNA of superior quality, which can be used directly for downstream sequencing applications.
This webinar focuses on the development of a semi-automated workflow to profile the gut microbiota of young and old individuals and identify changes in bacterial composition and function that occur with age. This workflow will help to simplify and streamline the DNA extraction process for samples with high inhibitor content and subsequent microbial community analyses.
Elisa - an introduction to the basic principles and assay formats presentationExpedeon
- Enzyme-Linked Immunosorbent Assay;
- Immunoassay utilising antibodies linked to enzymes for detection by colour change;
- Evolved from RIA in the 1960s;
- Antibody or analytebeing detected is absorbed to a solid surface, meaning unbound materials can be washed away with ease;
- With time, more techniques were developed and ELISA is now used to describe any assay where a molecule is absorbed on a solid phase.
it helps to known about ELISA ,it is one of the technique,, its type , its benefits & disadvantages , it also help to known about antigen,antidody, immunity etc
ELISA is a well know term that is an abbreviation of Enzyme Linked Immunosorbent Assay. This microplate based technique relies on the use of an antibody that has been linked to an enzyme. In the presence of an appropriate substrate, enzymatic activity produces a color change as the ELISA readout, which can be measured and provides information about the presence and quantity of the target antigen in the sample material.
Electrophoresis is a simple, rapid, and highly sensitive analytical technique to study the properties of proteins and nucleic acids, and has become a principle tool in analytical chemistry, biochemistry, and molecular biology. Polyacrylamide gel electrophoresis (PAGE) can be used to analyze the size, amount, purity, and isoelectric point of polypeptides and proteins. Sodium dodecyl sulfate polyacrylamide discontinuous gel electrophoresis (SDS PAGE) is the most commonly used system whereby proteins become separated strictly by their size, but there are different variations of this technique.
Antibody-oligonucleotide (Ab-Oligo) conjugates have been used in
numerous applications from diagnostics to therapeutics and were
developed through an unmet need for precise and efficient detection of low-abundance proteins. Ab-Oligo conjugates have since played a significant role in enhancing an extensive range of biological techniques that include immunological and proteomic research, biomarker discovery, clinical diagnostics – including point-of-care, as well as other novel techniques. Antibodies can be readily conjugated to oligonucleotides via their amino acid residues, making them suitable for most in vitro applications, as they possess several functional groups.
His Tag Protein Production and PurificationExpedeon
The study of protein regulation, structure, and function relies heavily on the expression and purification of recombinant proteins. Many recombinant proteins are expressed as fusion proteins, meaning that they contain an affinity / epitope tag. A tag is a short sequence of DNA that codes for a specific amino acid, which is frequently inserted into a target gene at the point of coding for expression at either the N or C terminal of the protein required.
GELFrEE® 8100 Fractionation System Tech NoteExpedeon
Successful sample preparation is a key step during any analytical
procedure and begins with a defined experimental design. Important steps in sample preparation include proteolytic digestion of proteins into peptide fragments, and peptide fractionation. This is especially important prior to applications such as mass spectrometry (MS).
Antibody-oligonucleotide (Ab-Oligo) conjugates have been used in
numerous applications from diagnostics to therapeutics and were
developed through an unmet need for precise and efficient detection of low-abundance proteins.
Proteomics of small proteins from plant tissuesExpedeon
Small genes and the proteins that they encode can play important biological roles including signaling, development, and mediation of plant-microbe interactions in organisms ranging from bacteria to plants to mammals (Frith et al.; Basrai et al.; Galindo et al.; Hemm et al. 2008, 2010; Kastenmeyer et al.). However, genes that encode proteins containing <100 residues are difficult to identify reliably solely by DNA sequence analysis (Dinger et al.)
Proteomic profiling of fractionated post-myocardial infarctionExpedeon
Acute myocardial infarction remains a leading cause of morbidity and mortality worldwide.Heart failure is the result of adverse remodeling of the collagenous scar that replaces the
damaged myocardium after MI. Markers of LV remodeling can be either identified in the circulation (e.g. serum or plasma) or detected in the heart by imaging technologies or biopsy.
NVoy technology is a quantum leap in protein processing, production and analysis. It uses proprietary NV polymers to enhance protein solubility and stability through the formation of multi-point reversible complexes with proteins without altering their structure.
Circular dichroism spectroscopy is an analytical technique used to estimate the secondary and tertiary structure of proteins. This technique can be used to confirm whether structure has been retained during protein processing, but is frequently adversely affected by additives such as solubility enhancers and detergents.
NVoy technology is a quantum leap in protein processing, production and analysis. It uses proprietary NV polymers to enhance protein solubility and stability through the formation of multi-point reversible complexes with proteins without altering their structure.
Protein processing and production is often hampered by the formation of aggregates that restrict and complicate
the handling of proteins, antibodies and enzymes. NVoy is designed to minimise the sequential losses in consecutive
protein processing steps which would otherwise dramatically reduce the overall protein yield.
NVoy technology is a quantum leap in protein processing, production and analysis. It uses proprietary NV polymers to enhance protein solubility and stability through the formation of multi-point reversible complexes with proteins without altering their structure.
NVoy technology is a quantum leap in protein processing, production and analysis. It uses proprietary NV polymers to enhance protein solubility and stability through the formation of multi-point reversible complexes with proteins without altering their structure.
Top down proteomics of soluble and integral membrane proteinsExpedeon
Mitochondria provide important cellular functions including
oxidative phosphorylation, fatty acid biosynthesis, and acting as
gatekeepers to apoptosis.
GELFrEE1 affords rapid mass-based protein separation over a range 10-150 kDa. Here, we demonstrate a multiplexed design enabling increased loading capacity and throughput. We
demonstrate comprehensive analysis of the yeast proteome using GELFrEE coupled to LC-MS/MS analysis.
Identification and characterization of intact proteins in complex mixturesExpedeon
The ability to fully characterize proteins in their intact forms allows thorough biological investigation of the functional importance of changes such as post-translational modifications, protein isoforms/sequence variations, and protease cleavages.
Improved coverage of the proteome using gel eluted liquidExpedeon
It has long been understood that sample fractionation is critically important to generating quality, comprehensive proteomics data. In spite of the continual improvements in speed and sensitivity of mass spectrometers, these instruments are still unable to adequately overcome the enormous challenge
of most biological samples without multiple dimensions of separation prior to mass analysis.
Optimization of experimental protocols for cellular lysisExpedeon
In this project, we have compared existing sample preparation methods for proteomics studies against newly developed FASP method and our in-house developed SDS-TCA protocol. For our
preliminary studies, we have chosen a very well characterized soil microbe Pseudomonas putida.
Characterization of intact antibodies by pre-fractionation using gel electrop...Expedeon
Antibodies represent an important class of proteins due to their central role in the immune response. Moreover, there is an increasing interest in the use of recombinant antibodies as novel drug therapies.
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.
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.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
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.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
3. www.innovabiosciences.com
Agenda
• ELISA and real time PCR - advantages, disadvantages
• Main steps of Immuno-PCR assay
• Introduction to Thunder-Link® PLUS oligo labeling technology
• Q&A session
6. www.innovabiosciences.com
What is immunodetection?
• Immunodetection is the process of using antibodies to detect
antigens in a sample
• Western blotting
• Immunocytochemistry
• Immunohistochemistry
• Flow cytometry
• ELISA
• Sensitive
• Selective
• Wide variety of analytes
7. www.innovabiosciences.com
What is immuno-PCR?
• Developed in 1992 by Sano et al
“Immuno-PCR: very sensitive antigen detection by means of specific antibody-DNA
conjugates” Science 258 (5079) 120-122
• Combines the antibody-based specificity of an ELISA with the
amplification of PCR
• Extremely high levels of sensitivity
8. www.innovabiosciences.com
ELISA
• Enzyme-Linked Immunosorbent Assay
• Evolved in the 1970s from radioimmunoassay (RIA)
• Relies on an antibody which has been linked to an enzyme
• Horseradish peroxidase, Alkaline Phosphatase, Glucose Oxidase
• Colorimetric readout
• Alternative readouts have been developed
• Fluorescence
• Luminescence
9. www.innovabiosciences.com
ELISA – advantages & disadvantages
Advantages Disadvantages
• Easy to perform
• Quantitative immunoassay
• Sensitive – limit of detection typically µg – pg
• Relatively inexpensive
• Can be used to screen large numbers of
samples
• Amenable to automation – suitable for High
Throughput Screening
• Cannot be used to measure parameters
such as tissue distribution or molecular
weight of the target
• The use of labeled secondary antibodies
for detection can introduce background
signal
• Some readouts are not amenable to
multiplexing
11. www.innovabiosciences.com
Lightning-Link® for direct antibody labeling
✓ Quick and easy to use
✓ Only 30 seconds hands-on time
✓ No separation steps - 100% recovery
✓ Scalable
Lightning-Link® labeling kits
• Enzymes – HRP, AP, Glucose oxidase
• Fluorescent dyes
• Fluorescent proteins
• Biotin
• Streptavidin
New product! Europium conjugation kit for covalent attachment of antibodies or proteins to
specially-treated 200nm europium particles – ideal for time resolved fluorescence (TRF)
13. www.innovabiosciences.com
PCR detection methods – gel electrophoresis
• Ethidium bromide was traditionally used to
visualize DNA following gel electrophoresis
• DNA interchelator
• Requires a UV transilluminator for visualization
• Molecular weight markers and restriction enzymes
can be used to confirm amplification of the target
• Southern blotting can provide additional
confirmation
DNA stained with EtBr
fluoresces under UV light
14. www.innovabiosciences.com
PCR detection methods – real time PCR/qPCR
• Various fluorescent chemistries have been designed to correlate
production of the PCR product to fluorescence intensity
• SYBR Green Dye
• Taqman® probes
15. www.innovabiosciences.com
Real time PCR – advantages & disadvantages
Advantages Disadvantages
• Rapid – no post-reaction processing is
necessary
• Increased sensitivity - can detect smaller
amounts of starting material than traditional
PCR
• Quantitative rather than qualitative
• Large dynamic range
• Significantly greater sample throughput
• Requires expensive instrumentation
16. www.innovabiosciences.com
Real time PCR – data analysis
• The PCR reaction can be divided in to four phases
• Linear ground phase – fluorescence is below
background levels, baseline fluorescence is calculated
• Early exponential phase – fluorescence crosses a
defined threshold, the cycle at which this occurs is
known as Ct
• Exponential phase – optimal amplification
• Plateau phase – reagents become limiting, the
reaction slows down
18. www.innovabiosciences.com
Real time PCR - instrumentation
• A wide range of instrumentation is commercially available
• Factors to consider when selecting a reader include:
• Price
• Size
• Block format
• Well capacity
• Individually programmed wells
• Heated lids
• Maximum ramp rate / run time
• Software integration......
19. www.innovabiosciences.com
Immuno-PCR
ELISA
• Adaptable to the detection of any protein
• Not adequate for the detection of low
abundance analytes
Real time PCR
• Provides exponential signal amplification
• Cannot be used directly for antigen
detection
Immuno-PCR
21. www.innovabiosciences.com
Immuno-PCR – assay optimization
• Maximize the specific readout, minimize background signal
• Plate coating
• Plate washing
• Blocking
• Sample incubation
• Plate layout
• Choice of antibodies
22. www.innovabiosciences.com
Immuno-PCR – plate coating
• Antibody binding via non-specific adsorption
• Typically performed overnight at 4oC
• Carbonate, bicarbonate or borate buffer at pH>9
• Optimal concentration of the coating antibody should be
determined
• Starting range of 0.2 - 20µg/ml
• Grid layout for simultaneous testing of multiple antibody
concentrations
23. www.innovabiosciences.com
Immuno-PCR – plate washing
• Wash steps are essential for the removal of unbound
reagents
• Typical wash buffers include Phosphate Buffered Saline
(PBS) and Tris Buffered Saline (TBS)
• A low concentration of detergent (0.01-0.1% v/v) is often
added
• The number and duration of washes should be optimized
24. www.innovabiosciences.com
Immuno-PCR – blocking
• Blocking is critical to prevent non-specific binding
• Maximizes signal: background (S:B) ratio
• The blocking buffer is typically a weak solution (1%-5%
w/v) of protein diluted in wash buffer
• Bovine Serum Albumin (BSA)
• Non-fat dried milk
• Gelatin
• Casein
• Whole serum
• An overnight blocking step can improve S:B
25. www.innovabiosciences.com
Immuno-PCR – sample incubation
• Dilution of test samples can reduce background effects
caused by the nature of the sample material
• Short incubation times are often sufficient for antibody:
antigen binding
• Incubation temperature should be optimized
26. www.innovabiosciences.com
Immuno-PCR – plate layout
• Positive controls – spiked with known concentrations of antigen
• Negative controls – sample material without antigen
• Calibration curve
• Used to calculate the antigen concentration of test samples
• 10-fold serial dilutions, or lower, of the antigen
• All samples should be analyzed in duplicate
• Controls and standards should be diluted in the same matrix as the
test samples
27. www.innovabiosciences.com
Immuno-PCR – choice of antibodies
• Requires highly specific antibodies
• Recognition of a common molecular motif will increase
background signal
• Capture and detection antibodies should each target a
different epitope
Polyclonal Monoclonal
• Subject to variability between different
immunized animals
• A permanent antibody supply cannot be
guaranteed
• Immunogen affinity purification is
required
• Large quantities of a specific antibody,
recognizing a single antigenic epitope
• Uniform performance
• Immunogen affinity purification is not
necessary
28. www.innovabiosciences.com
Immuno-PCR – advantages & disadvantages
Advantages Disadvantages
• Adaptable to the detection of any protein
• Provides exponential signal amplification
• Extremely low limit of detection (pg - fg)
• Suitable for small sample volumes
• Compatible with complex samples
• Fewer incubation steps than an ELISA, improved
assay reproducibility
• Rapid time to results
• Wider dynamic range than an ELISA
• Amenable to multiplexing
• Requires conjugation of an
antibody to an oligonucleotide
29. www.innovabiosciences.com
ThunderLink® PLUS
• An easy-to-use kit for the production of
stable antibody-oligonucleotide conjugates
• No specialist knowledge required
• 30 minutes activation, 60 minutes
conjugation
• Optional clean-up step to remove unbound
oligo
30. www.innovabiosciences.com
ThunderLink® PLUS - advantages
Advantages
• Quick and easy to use
• High levels of antibody and oligo recovery
• Covalent bond
• Unidirectional chemistry
• Use your own oligo and antibody, at your desired ratio
• Linking chemistry works at both the 5’ and the 3’ end
• Optional post-conjugation clean-up step for removal of unbound oligo
• Positive control antibody and oligo included to confirm conjugation success
• Freeze dried
• Stringently QC tested
31. www.innovabiosciences.com
ThunderLink® PLUS – antibody considerations
• Activation of 100µg antibody in 100µl suitable
buffer
• Purified antibody, 1mg/ml
• AbSelect™ antibody purification kits
• Buffer exchange
• Concentration of the antibody
• Purification from TCS, ascites or serum
Buffer components Antibody buffer
pH 7-9
Amine free buffer ✓
Non-buffering salts (e.g. NaCl) ✓
Chelating agents (e.g. EDTA) ✓
Sugars ✓
Glycerol <50%
Thimerosal
Merthiolate
Sodium azide* <0.1%
BSA* <0.1%
Gelatin* <0.1%
Tris <20mM
Glycine
Primary amines (e.g. amino acids)
Thiols (e.g. mercaptoethanol, DTT)
* Individually the concentrations shown should not affect the reaction, however
in combination with additional compounds that are not recommended above a
certain concentration the reaction may be affected.
32. www.innovabiosciences.com
ThunderLink® PLUS – oligo considerations
• Activation of 60-100µM oligo in 100µl suitable
buffer
• Must be HPLC purified
• Single-stranded oligos of 10-120 bases
• Terminal amine group
• Double-stranded oligos up to 80 base pairs
• Only one strand should be terminally aminated
• Linking chemistry works at either end of the
oligo
• Compatible with biotinylated or fluorescent dye-
conjugated oligos
Buffer components Oligonucleotide buffer
pH 6-8
Amine free buffer ✓
Non-buffering salts (e.g. NaCl) ✓
Chelating agents (e.g. EDTA) ✓
Sugars ✓
Glycerol <50%
Thimerosal
Merthiolate
Sodium azide* <0.1%
BSA* <0.1%
Gelatin* <0.1%
Tris
Glycine
Primary amines (e.g. amino acids)
Thiols (e.g. mercaptoethanol, DTT)
* Individually the concentrations shown should not affect the reaction, however
in combination with additional compounds that are not recommended above a
certain concentration the reaction may be affected.
33. www.innovabiosciences.com
ThunderLink® PLUS – antibody: oligo ratio
• The antibody: oligo ratio can easily be varied
• The antibody: oligo ratio is an average since a population of labeled antibodies will
be produced
• The conjugation efficiency is typically 95-100%
Volume of activated
antibody (µl)
Volume of activated
oligo (µl)
Volume of wash buffer
(µl)
Antibody: oligo ratio
300 300 0 1:15
300 200 100 1:10
300 100 200 1:5
300 60 240 1:3
300 20 280 1:1
34. www.innovabiosciences.com
Custom services
• Antibody-oligo micro-optimization service
• Six conjugates – different chemistries, different antibody: oligo ratios
• Custom antibody-oligo conjugation service
• Production of chosen conjugate at required scale
• Both services include
• Antibody clean-up prior to conjugation
• Removal of unbound oligo post-conjugation
• QC gel to demonstrate successful conjugation
35. www.innovabiosciences.com
Summary
• Immuno-PCR combines the specificity of an ELISA with the sensitivity
of PCR
• Adaptable to the detection of any protein
• Rapid generation of robust and reproducible data
• Amenable to multiplexing
• Thunder-Link® PLUS for quick and easy conjugation of antibodies to
oligonucleotides
Thank you!
38. www.innovabiosciences.com
Get in touch..
For more information please get in touch or visit our website
www.innovabiosciences.com
+44 (0) 1223 661000
www.linkedin.com/company/innova-biosciences@InnovaBioSciwww.facebook.com/InnovaBiosciences