overview document showing how scientist can easily and quickly use Anteo's Mix&Go surface coating to attach biomolecules, e.g. antibodies, streptavidin, Protein A to a variety of synthetic surfaces from nanobeads to glass slides withut causing damage to the biomolecule
Medical Research: conflicts between autonomy and beneficence/non maleficence, euthanasia, informed consent, confidentiality, criticisms of orthodox medical ethics
Prepared By :
Subrata SInha (Senior Analyst)
Analytical method validation’s superlative characteristic, linearity is comprehensively explained in this slide show. Linearity commonly understood as proportional relationship between two units, in context of method validation, it refers toproportional relationship between test results and concentrations.
Medical Research: conflicts between autonomy and beneficence/non maleficence, euthanasia, informed consent, confidentiality, criticisms of orthodox medical ethics
Prepared By :
Subrata SInha (Senior Analyst)
Analytical method validation’s superlative characteristic, linearity is comprehensively explained in this slide show. Linearity commonly understood as proportional relationship between two units, in context of method validation, it refers toproportional relationship between test results and concentrations.
Paper chromatography is an analytical method used to separate coloured chemicals or substances.It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC).
ACRI is a leading clinical research training institute in Bangalore.
ACRI creates a value add for every degree. Our PGDCRCDM course is approved by the Mysore University. Graduates and Post Graduates and even PhDs have trained with us and got enviable positions in the Clinical Research Industry. ACRI supplements University training with Industry based training, coupled with hands-on internships and projects based on real case studies. The ACRI brand gives the individual the confidence and expertise to join the ever-growing workforce both in the country and abroad.
Gas chromatography is one of the widely used chromatographic techniques, which use inert gas as the mobile phase.
In gas chromatography the components of a sample, after vaporization, are separated by being partitioned between gaseous mobile phase and solid or liquid stationary phase.
The inert gas does not interfere with the analyte but transport the components through the column and facilitate the separation.
The mobile phase is comprised of an inert gas such as helium, argon or nitrogen.
The stationary phase consists of a packed column in which the packing or solid support, or a liquid coat act as stationary phase.
The main principles involved are adsorption and partition for gas solid chromatography and gas liquid chromatography, respectively.
(I) MEDICAL RESEARCH_ UNIT_III_RESEARCH METHODOLOGY & BIOSTATISTICS.pptxRAHUL PAL
Research Methodology and Biostatistics syllabus:
Medical Research: History, values in medical ethics, autonomy, beneficence, non-maleficence, double effect, conflicts between autonomy.
Medical research has a long and varied history. It has evolved from rudimentary practices to sophisticated, evidence-based methodologies. Some key milestones include the development of the scientific method, the use of randomized controlled trials, the discovery of antibiotics, and the mapping of the human genome. Ethical concerns have also played a significant role in shaping the history of medical research, especially in response to various ethical violations, such as the Tuskegee Syphilis Study and the Nuremberg Trials.
Resolving conflicts between these principles often requires careful consideration, ethical analysis, and, in some cases, consultation with ethics committees or boards. The specific course of action may vary based on the individual circumstances and ethical frameworks employed by healthcare professionals and researchers. Ethical guidelines and regulations also play a significant role in addressing and preventing these conflicts in medical research.
FOMAT Medical Research is a site research network specializes in developing clinical. We offer a wide range of solutions for Sponsors, Clinical Contract Organizations (CROs), and Sites throughout the Americas. Visit here- https://www.fomatmedical.com
Criticisms of orthodox medical ethics, importance ofsupriyawable1
ethics is a very large and complex field of study with many branches .medical ethics is the branch of ethics that deals moral issues in medical practice. principles of medical ethics - autonomy ,beneficence ,confidentiality,do not harm,equity .importance of communication .
Drug regulatory agencies play a critical role in ensuring the safety, efficacy, and quality of pharmaceutical products, including drugs, biologics, and medical devices. These agencies are responsible for establishing and enforcing regulations and guidelines that govern the development, testing, approval, manufacturing, labeling, marketing, and post-market surveillance of these products.
Simple methods for forming protein monolayers lab on-a-chip 2013AnteoDx
Poster shown at the 2013 Lab-on-a-Chip conference showing experimental data created with the Mix&Go surface chemistry on various surfaces incl. glass, polystyrene and gold as well as comparisons to the standard methods.
Paper chromatography is an analytical method used to separate coloured chemicals or substances.It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC).
ACRI is a leading clinical research training institute in Bangalore.
ACRI creates a value add for every degree. Our PGDCRCDM course is approved by the Mysore University. Graduates and Post Graduates and even PhDs have trained with us and got enviable positions in the Clinical Research Industry. ACRI supplements University training with Industry based training, coupled with hands-on internships and projects based on real case studies. The ACRI brand gives the individual the confidence and expertise to join the ever-growing workforce both in the country and abroad.
Gas chromatography is one of the widely used chromatographic techniques, which use inert gas as the mobile phase.
In gas chromatography the components of a sample, after vaporization, are separated by being partitioned between gaseous mobile phase and solid or liquid stationary phase.
The inert gas does not interfere with the analyte but transport the components through the column and facilitate the separation.
The mobile phase is comprised of an inert gas such as helium, argon or nitrogen.
The stationary phase consists of a packed column in which the packing or solid support, or a liquid coat act as stationary phase.
The main principles involved are adsorption and partition for gas solid chromatography and gas liquid chromatography, respectively.
(I) MEDICAL RESEARCH_ UNIT_III_RESEARCH METHODOLOGY & BIOSTATISTICS.pptxRAHUL PAL
Research Methodology and Biostatistics syllabus:
Medical Research: History, values in medical ethics, autonomy, beneficence, non-maleficence, double effect, conflicts between autonomy.
Medical research has a long and varied history. It has evolved from rudimentary practices to sophisticated, evidence-based methodologies. Some key milestones include the development of the scientific method, the use of randomized controlled trials, the discovery of antibiotics, and the mapping of the human genome. Ethical concerns have also played a significant role in shaping the history of medical research, especially in response to various ethical violations, such as the Tuskegee Syphilis Study and the Nuremberg Trials.
Resolving conflicts between these principles often requires careful consideration, ethical analysis, and, in some cases, consultation with ethics committees or boards. The specific course of action may vary based on the individual circumstances and ethical frameworks employed by healthcare professionals and researchers. Ethical guidelines and regulations also play a significant role in addressing and preventing these conflicts in medical research.
FOMAT Medical Research is a site research network specializes in developing clinical. We offer a wide range of solutions for Sponsors, Clinical Contract Organizations (CROs), and Sites throughout the Americas. Visit here- https://www.fomatmedical.com
Criticisms of orthodox medical ethics, importance ofsupriyawable1
ethics is a very large and complex field of study with many branches .medical ethics is the branch of ethics that deals moral issues in medical practice. principles of medical ethics - autonomy ,beneficence ,confidentiality,do not harm,equity .importance of communication .
Drug regulatory agencies play a critical role in ensuring the safety, efficacy, and quality of pharmaceutical products, including drugs, biologics, and medical devices. These agencies are responsible for establishing and enforcing regulations and guidelines that govern the development, testing, approval, manufacturing, labeling, marketing, and post-market surveillance of these products.
Simple methods for forming protein monolayers lab on-a-chip 2013AnteoDx
Poster shown at the 2013 Lab-on-a-Chip conference showing experimental data created with the Mix&Go surface chemistry on various surfaces incl. glass, polystyrene and gold as well as comparisons to the standard methods.
Radiation-Cured Components & Their Use in Hard, Scratch Resistant Coating App...Sartomer
This presentation covers the following topics:
- Current and potential hard coat applications
- Description of products tested and their attributes
- Taber haze and Taber abrasion resistance
- Weathering resistance
- Barrier properties of a typical hard coat formulation
- Polyurethane dispersion products (PUDs)
You can visit Sartomer at Sartomer.com and follow them on Twitter @SartomerGlobal and on LinkedIn.
Protein binding and non binding surfaces on biochips lab on chip 2012AnteoDx
Poster presented by Anteo Diagnostics at the Lab-on-a-chip 2012 conference. The posters shows data about how to create regions that bind and regions that do not bind proteins on a variety of different surfaces using the Mix&Go surface chemistry. The ability to create binding and non-binding regions is crucial in many point-of-care, IVD and lab-on-a-chip applications.
OECD Webinar | Assessing the dispersion stability and dissolution (rate) of n...OECD Environment
On Thursday 25 February 2021, Anne Gourmelon (Environment Directorate, OECD), Kathrin Schwirn (German Environment Agency, Umweltbundesamt, UBA); Frank von der Kammer (University of Vienna) Research and Development Center) and Doris Völker (German Environment Agency, Umweltbundesamt, UBA) presented the scope, content, and use of the Test Guideline No. 318: Dispersion Stability of Nanomaterials in Simulated Environmental Media and its accompanying Guidance Document. Further discussions focused on the scope of the upcoming Test Guideline.
The increased production and wide usage of manufactured nanomaterials suggest a higher probability of finding them in the environment. Therefore, testing the dissolution rate and dispersion stability for toxicity assessment are of paramount importance for adequate hazard assessment.
Experimental Characterization of Carbon Fibre T700 / Epoxy towpreg for Space ...IJERA Editor
This document covers detailed experimental characterization of Carbon Fibre T 700/Epoxy towpreg.The
experimental characterization of carbon fibre T 700/Epoxy towpreg composite material is necessary required for
generation of mechanical properties data for analysis, design, and fabrication of structural components using
that material and for quality control of the material. The testing of composite materials offers unique surprises
because of the special characteristics of composites. Factors not considered important in metals testing are very
important in testing composites. For example, composites are anisotropic, with properties that depend on the
direction in which they are tested. Speed must be carefully monitored at the time of testing of specimens and
also fiber content, void content, specimen conditioning (drying, storage, etc) have important effects on material
properties.
In order to design composite products, a thorough experimental characterization of carbon fibre T 700 / Epoxy
towpreg composite material and its behaviour is necessary.
Similar to Mix&Go Surface Chemistry - How to Easily Attach a Protein to a Synthetic Surface (20)
Basavarajeeyam is an important text for ayurvedic physician belonging to andhra pradehs. It is a popular compendium in various parts of our country as well as in andhra pradesh. The content of the text was presented in sanskrit and telugu language (Bilingual). One of the most famous book in ayurvedic pharmaceutics and therapeutics. This book contains 25 chapters called as prakaranas. Many rasaoushadis were explained, pioneer of dhatu druti, nadi pareeksha, mutra pareeksha etc. Belongs to the period of 15-16 century. New diseases like upadamsha, phiranga rogas are explained.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
2. Developing a Universal “Glue”
• What is Mix&Go?
• aqueous metal polymers which use multiple weak binding
forces to form very strong bonds.
• Mix&Go surfaces are;
• stable for years but in the presence of proteins rapidly bind
with minimal conformational damage.
• acts as a molecular “velcro” for proteins, synthetic polymers,
nanoparticles, etc.
• Characteristics of proteins bound to Mix&Go surfaces are:
• functionally active protein mono-layers
•
Resulting benefits of Mix&Go are:
• many and varied
3. What Is Mix&Go?
A family of aqueous metal polymers bind to virtually all
surfaces used in life sciences
Configurations
M
M
M
M
M
• Metal oligomers with different
shape and charge densities
• Determines speed and
strength of binding to surfaces
and biomolecules, e.g. proteins
Mix&Go is based on Cr(III)
• Slow exchange half life; order of hours.
• Not considered a health risk from numerous studies, e.g.,
International Agency for Research on Cancer (IARC)
• Not considered hazardous waste by US EPA
• An essential elements in human health
4. Forming Mix&Go Activated Surfaces
•
•
Mix&Go is a cationic polymer: mechanism of binding is via both
electrostatic and co-ordination forces
Making Mix&Go surfaces is easy, e.g.:
Microtiter plates:
Pipette 100 L
Mix&Go into
wells, incubate
for 30 minutes
and wash
•
Beads:
Mix&Go
Add
nanoparticles
Mix well
Activated in 30
mins and stable
for years
Mix&Go is reactive with:
• Acid, amine, hydroxy, epoxy and other polymer beads
• Nanoparticles: gold colloids, iron oxides, etc.
• Sepharose, poly(vinylalcohol), poly(hydroxyethylmethacrylate)
• Glass, silica oxides, titanium oxides, ceramics, etc.
• Polystyrene microtitre plates
• Engineering thermoplastics such as COC/COP
5. Increased Protein Stability
Streptavidin microarray slides need to be stored at -20°C and have shelf
life of hours at RT. Streptavidin on Mix&Go coated microarray slides
show excellent stability even after 14 days at 25 C and 37 C.
18,000
16,000
14,000
12,000
10,000
8,000
6,000
4,000
2,000
0
Freshly Coated
200 μg/mL
50 μg/mL
10 μg/mL
18,000
16,000
14,000
12,000
10,000
8,000
6,000
4,000
2,000
0
14 Days @ 25 C
15,000
14 Days @ 37 C
10,000
5,000
0
200 μg/mL
50 μg/mL
10 μg/mL
200 μg/mL
Stability studies using binding capacity of biotinylated mouse
Ab shows no difference even after 2 weeks at 37 C.
50 μg/mL
10 μg/mL
6. Streptavidin on Mix&Go Microarrays
Testing existing commercial products. Biotin-RPE binding on Streptavidin
Mix&Go microarray slides vs commercial standard (3D Type).
Top: Biotin-RPE binding on
Streptavidin Mix&Go slides
(far left) vs commercial
product (left).
12.5 25 40 100 200
12.5 25 40 100 200
g/ml biotin-RPE
g/ml biotin-RPE
12.5 25 40 100 200
g/ml biotin-RPE
Bottom: Same experiment
with prior biotin blocking
shows no binding on Mix&Go
slides (far left). Commercial
product binding is all Non
Specific Binding (NSB)
(left).
12.5 25 40 100 200
g/ml biotin-RPE
7. Streptavidin on Mix&Go on Plates
Streptavidin Mix&Go plates (different brands) compared to Thermo/Pierce
Streptavidin plates. Streptavidin coating at 2 g/ml, 30 mins.
2.500
Mix&Go on
Greiner Low
Bind Block
2.000
OD (450-620nm)
Mix&Go on
Nunc Polysorp
Block
1.500
Mix&Go on
Corning Med
Bind Stripwell
1.000
Thermo High
Sensitivity
Stripwell
0.500
Thermo
Standard Block
0.000
0
100
200
300
400
biotin Mouse Concentration (ng/mL)
500
Using only 200 ng/well and 30 min coating time,
streptavidin coated onto Mix&Go plates perform as well or
better than existing commercial products.
600
8. Decreased Protein Usage (on Plates)
GM-CSF cAb titration on Mix&Go activated Greiner low binding plates
vs. passive binding on Nunc Maxisorp plates
2.5
Mix&Go
cAb Conc
OD (450-620nm)
1.0 g/mL
$1.10
$8.80
Plate Type
Greiner
Low Bind
Nunc
Maxisorp
Plate Cost*
1.5
0.125 g/mL
cAb Cost
per Plate*
2
Passive
$0.55
$4.92
1
Mix&Go
Greiner
Passive Nunc
Maxisorp
0.5
0
0
2
4
6
cAb (ug/mL)
8
10
The binding kinetics of Mix&Go activated polystyrene surface
results in peak signal at lower antibody concentration and plateaus
regardless of increased concentration.
Significant COGS savings can be achieved with Mix&Go
9. Improving Antibody Distribution
Intra-plate CVs on TNFα sandwich assay. Comparison of
Mix&Go activation vs passive on Greiner Low Binding plates
Mix&Go
Passive
Greiner Low Bind Plate Passive
Greiner Low Bind Plate - Mix&Go
1.2-1.4
0.4-0.6
1-1.2
0.2-0.4
0.8-1
A
D
1.4
1.2
1
0.8
12
10 11
G
8 9
6 7
4 5
3
CV = 4.43%
1 2
A
0.6
0.4
0.2
D
11 12
9 10
G
7 8
5 6
3 4
1 2
Mix&Go activation decreases well-to-well variation
while increasing activity compared to passive binding
on Greiner low binding plates
CV = 9.21%
10. Stability on Plates
Change in activity from Day 0
IL-6 Stability Testing
1.800
Temp = 37 C
Mix&Go
Passive
Day 0 vs.
Day 8
1.17%
8.64%
Day 0 vs.
Day 15
7.55%
17.75%
1.600
1.400
Day 0
OD (450-620nm)
1.200
1.000
Day 8 37C
0.800
Day 15 37C
0.600
0.400
0.200
0.000
Mix&Go on
Greiner Medium
Stripwell
Passive on
Greiner High
Bind Stripwell
Accelerated stability @ 15 days
stored at 37 C equates to
>1year stability stored at 4 C
Mix&Go C10 activated plates
coated with IL-6 and blocked
with 10mM PBS + 1% BSA +
5% Sucrose
Mix&Go plates maintain protein stability better than passive binding.
11. Advantages w/ Decreasing Surface
Comparison of 96 well (Left) and 384 well (Right) micro-titre plates. TNFα
sandwich assay using cAb conc of 0.5 g/ml
3.000
3.000
2.500
2.500
2.000
2.000
1.500
1.500
1.000
1.000
OD (450-620nm)
Mix&Go
No Mix&Go
0.500
0.500
0.000
0.000
0
1,500 3,000 4,500 6,000 7,500 9,00010,500
Ag Concentration (pg/mL)
0
1,500 3,000 4,500 6,000 7,500 9,000 10,500
Ag Concentration (pg/mL)
Antibody conformation and orientation are preserved resulting in more
functional antibodies. The effect is more pronounced on smaller surface
areas where each antibody becomes more important.
12. Coating on Other COC/COP Plastics
Loading assay using capAb (1 g/ml) and detection using GAM-HRP
(0.1 g/ml) on passively bound vs. Mix&G activated COC surfaces.
24 hours
TNFa
1.0
3
O.D. (450 nm)
O.D.
2.5
0.5
0.0
2
TNF-a
1.5
TnI
TSH
1
0.5
0
20
40
60
Time (min)
80
100
0
COC
M&GC10
Capture antibody binding efficiency to Mix&Go activated COC
surfaces is far faster than passive binding. Even after 24 hr
coating, passive binding did not reach Mix&Go levels.
13. Gold Colloids on Mix&Go COC Surfaces
Mix&Go can bind more than just proteins. AFM images of gold colloids
(40 nm) on COC and Mix&Go activated COC surfaces.
Gold nanoparticles on Mix&Go
activated COC
Gold nanoparticles on COC
by passive binding
5
5
4
10
6
4
4
5
2
µm
0
nm
µm
3
2
0
2
-2
-5
1
-4
1
-10
-6
0
0
1
2
3
µm
4
5
0
0
1
2
3
4
5
µm
Surface coverage of gold colloids on COC by passive binding
was calculated to be 11%. Mix&Go COC surfaces gave 64%.
nm
3
14. Sandwich Assays on Magnetic Particles
IFN-ϒ chemiluminescense assay on Mix&Go MyOne (Life Technologies)
vs. MyOne Tosyl magnetic particles.
Loading Assay: Mix&Go vs. Tosyl Beads
160000
140000
180,000
Antibody per mg beads
Mix&Go: 15ug Ab
Tosyl: 40ug Ab
120000
15ugAb+15ugBSA/mg MyOne Mix&Go beads
160,000
40ugAb/mg MyOne Tosyl beads
140,000
120,000
RLU
RLU (Goat anti Mouse IgG - HRP)
180000
Sensitivity Assay: Mix&Go vs. Tosyl Beads
100000
100,000
80000
80,000
60000
60,000
40000
40,000
20000
20,000
Signal to Noise (S/N)
Mix&Go: 2607
Tosyl: 1787
-
0
MyOne Mix&Go
Dynabeads
MyOne Tosyl Dynabeads
0
500
1000
1500
IFN-ϒ antigen pg/mL
2000
S/N of Mix&Go MyOne beads is superior to MyOne Tosyl beads using
less than half the amount of capture antibody. No co-coupling reagent
such as BSA were used.
2500
15. Bead Handling Characteristics
Mix&Go MyOne vs. Tosylated MyOne*
Left two tubes: Antibody Mix&Go
MyOne Dynabeads
Right two tubes: Antibody MyOne
Tosyl Dynabeads
Left two tubes: Antibody Mix&Go M270 Dynabeads
Right two tubes: Antibody M-280
Tosyl Dynabeads
Mix&Go beads form cleaner bead plugs under a magnetic field than
Tosyl beads.
*Tosyl beads coupled at recommended concentrations
16. Streptavidin on 200 nm Latex Nanoparticles
18,000,000
30,000
16,000,000
14,000,000
12,000,000
Batch 1
10,000,000
Batch 2
8,000,000
6,000,000
4,000,000
2,000,000
RLU (Goat anti Mouse IgG - HRP)
RLU (Biotinylated Mouse IgG/GAMHRP)
Comparison of Streptavidin on 243 nm Mix&Go activated particles
with Thermo’s Power-Bind Streptavidin 294 nm particles
Batch 1
25,000
Batch 2
20,000
15,000
10,000
5,000
0
Streptavidin Mix&Go 243nm
Particles
Commercial Streptavidin Thermo
Power-Bind (294nm) Particles
Specific Binding
Biotinylated Mouse IgG/Goat anti Mouse
IgG–HRP
Streptavidin Mix&Go 243nm
Particles
Commercial Streptavidin Thermo
Power-Bind (294nm) Particles
Non Specific Binding
Goat anti Mouse IgG–HRP
Streptavidin on Mix&Go activated latex nanoparticles shows over 5x increase
in biotinylated antibody loading factoring in particle size.
17. Maintaining Brownian Motion
Antibody on Mix&Go coated magnetic nanoparticles (500 nm).
Brownian motion under magnetic field in pooled plasma (TnI
assay) http://youtu.be/UB7mctqmP-c
Mix&Go activated Ademtech
beads coupled with 560 mAb
(125ug Ab/mg beads) in
pooled plasma
Colloidal stability of nanoparticles after Mix&Go activation and
protein coupling is maintained with particles below 40 nm.
18. Mix&Go on Gold Colloids
Localized Surface Plasmon Resonance (LSPR) studies on gold colloids
(Sigma, 100 nm). Mix&Go forms a thin film (<1 nm) on which binds a
mono-layer of proteins.
OD Normalized
Mix&Go
400
500
600
700
Mix&Go +
IgG
Passive +
IgG
OD Normalized
Passive
800
Wavelength (nm)
Mix&Go coating resulted in a LSPR
λ-max shift of 4nm with improved
colloid dispersity.
400
500
600
Wavelength (nm)
700
800
On Ab addition, there is a further λ-max shift
of 9nm on Mix&Go gold colloids. Without,
Mix&Go, the colloids aggregate.
19. Improving the Flow of Nanoparticles
Mix&Go
No Mix&Go
Particle
Abs
TEM analysis of Ab bound to
Mix&Go iron oxide nanoparticles
(10-15 nm).
The hydrophilic nature of Mix&Go particles allow better “flow” in
aqueous environments giving faster pull down speeds under magnetic
field.
20. Antibody on Mix&Go Magnetite
45,000,000
Two step functionalisation of super
paramagnetic nanoparticles;
Mix&Go addition, wash than
antibody addition.
41,501,039
40,000,000
35,000,000
RLU (GAM-HRP)
30,000,000
Chemiluminescent loading assay.
Detection with GAM-HRP (@
0.005ug/ml). Lumigen PS-atto was
used as the substrate.
25,000,000
20,000,000
The very large surface area of
magnetite nanoparticles give very
large protein binding capacity
giving high Signal to Background
of 3,106:1
15,000,000
10,000,000
5,000,000
13,360
Specific Signal
Background
Assay Readout
ESC-130416-1
21. Benefits of Mix&Go
• Forms stable activated surface on planer surfaces of glass, ceramics
or engineering plastics as well as beads and nanoparticles.
Benefits:
• universal, extremely stable coating for almost all surfaces
• water based chemistry is easy to use and manufacture
• 1 nm coating does not interfere with detection
• Forms undamaged protein mono-layer.
Benefits:
• Less non-specific binding
• Increased performance/sensitivity, esp. with limited surface area
• No need for anti-species Abs or Streptavidin constructs to bind
antibodies onto surfaces.
• Less protein and/or beads required.
Benefits:
• Reduced COGS
• Saves valuable/rare antibodies
• Improved reproducibility bead-to-bead, well-to-well, batch-to-batch.
Benefit:
• Improved accuracy with improved %CV.
22. Contact Information
Thank you very much for your interest in Mix&Go.
For more information please contact Anteo directly:
Headquarters
Anteo Diagnostics Ltd
Suite 4, 26 Brandl Street
Brisbane Technology Park
Eight Mile Plains, QLD 4113
Australia
Geoff Cumming, CEO
Geoff.cumming@anteodx.com
P:+61-417-203-021
US Contact
Tina Baumgartner
Tina.baumgartner@anteodx.com
P: +1-510-508-8462