Category
- Microscopy and Imaging » Confocal Microscopy
Booking Details
Facility Management Team and Location
samirmaji@iitb.ac.in
+(91-22) 2576 7774
- Prof. Sanjeeva Srivastava
- Prof. Ashutosh Kumar
- Prof. Samir K. Maji
- Prof. Kiran Kondabagil
- Prof. Sarika Mehra
- Prof. Ruchi Anand
Facility Features, Working Principle and Specifications
Surface plasmon resonance (SPR) is a label-free, real- time technique capable of measuring binding affinities and kinetics for bio-molecular interactions. SPR has become a key bio-sensing technology in the areas of biological research and medical sciences..
Specifications/Features
Can study interactions at physiological temperatures. Analysis temperature range 4°C to 45°C.
- Rapid buffer scouting application for fast assay development.
- Integrated buffer degasser ensures data quality at elevated temperatures.
- Designed to support large-scale research applications.
Special Features
- Designed to support large-scale research applications
- Supports 96- and 384-well microplates
- Up to 48 hours unattended operation
- Temperature control of sample compartment
- Analyze up to 384 samples per run
- Study interactions at physiological temperatures
- Analysis temperature range 4°C to 45°C
- Integrated buffer degasser ensures data quality at elevated temperatures
- Rapid buffer scouting for fast assay development
- Built-in buffer selector enables up to four different buffers to be tested at one time
- Sample recovery for identification by mass spectrometry
- Predefined software templates define entire recovery process
- Analytes recovered in a small volume, with minimal carry-over
- Deposition in vial containing digestion solution
- Software solutions for fast assay development, analysis, and evaluation
- High level of guidance for development and setup of assays
- Dedicated software support for data evaluation
Working Principle
Surface plasmon resonance (SPR) based instruments use an optical method to measure the changes in refractive index within about 150 nm from the sensor surface. They monitor molecular interactions in real time, using a non-invasive label-free technology that responds to changes in the concentration of molecules at a sensor surface as molecules bind to or dissociate from the surface. The essential components of a Biacore analytical system are sensor chip, optical detector and integrated microfluidic cartridge (IFC) (Figure 1). To study the interaction between two binding partners, one partner is attached to the sensor surface (ligand) and the other is passed over the surface through flow cells in sample solution (analyte). As the analyte binds to the ligand the accumulation of protein on the sensor surface causes an increase in refractive index. A sensorgram is a plot of response against time, showing the progress of the interaction (Figure 2). The SPR response is directly proportional to the change in mass concentration close to the surface.
Technical Specifications
- Detection technology: Surface Plasmon Resonance (SPR) biosensor
- Information provided: Kinetic and affinity data (KD, ka, and kd), specificity, selectivity, concentration, and thermodynamic data
- Data presentation: Result tables, result plots, and real-time monitoring of sensorgrams
- Analysis time per cycle: Typically 2 to 15 min
- Automation: 48 h unattended operation
- Sample type: LMW drug candidates to high molecular weight proteins (also DNA, RNA, polysaccharides, lipids, cells, and viruses) in various sample environments (e.g., in DMSO-containing buffers, plasma, and serum)
- Required sample injection volume: Plus 20 - 50 µl volume (application dependent)
- Injection volume: 2 to 350 µl
- Flow rate range: From 1 to 100 µl/min
- Flow cell volume: 0.06 µl
- Flow cell height: 40 µm
- Sample/reagent capacity: 1 x 96-or 384-well microplate and up to 33 reagent vials
- Analysis temperature: 4°C to 45°C (maximum 20°C below ambient temperature)
- Sample storage: 4°C to 45°C (maximum 15°C below ambient temperature)
- Sample refractive index range: 1.33 to 1.40
- Buffer selector: Automatic switching between 4 buffers
- In-line reference subtraction: Automatic
Instructions for Registration, Sample Preparation, User Instructions, Precautionary Measures and Charges
- Only online sample registration through the IRCC webpage Drona will be accepted.
- The user will be informed first about the sample run charges>after charges approval from PI< meeting date and time is given and later about the experiment date through drona.
- If the meeting/experimental appointment is made but the user can't go, kindly send an email immediately to spr.bios@iitb.ac.in to cancel the slot.
- Please come prepared with some literature reference papers describing as much as possible for sample preparation and experimental conditions for similar kind of studies.
- User should submit the samples along with the registration form.
- Experiments should be discussed with PI/TA and the facility in-charge before proceeding.
- Purity of samples is extremely important for generating good data.
- Protein concentrations should be measured accurately before starting the experiment.
- The molecular weight as well as the pI of the proteins should be known before immobilization.
- All buffers should be filtered through 0.22 μm filters and degassed.
- Do not degas buffers containing detergent. Add detergent after degassing.
- For organic solvent containing buffer, filter using organic solvent resistant membrane.
- Cell extracts and nanoparticles can block integrated micro fluidic cartridges and syringes.
- Any query regarding your SPR experiment can be emailed on spr.bios@iitb.ac.in
- Appointments will be provided as per queue and the user will be informed about the same.
- Kindly perform some literature review on this kind of work performed on either same or similar samples. Accumulate as much information as possible for better quality results.
Applications
Assay Development for binding studies for Biomolecule,Small molecules,Drugs binding studies.
Binding study of these molecules in crude lysates,whole cells ,etc
Interaction Kinetics analysis of Proteins,Nucleic acid,carbohydrates,Lipids,Small molecule,Drugs.
Mab screening.
Epitope Binning.
Immunogenicity testing
Calibration Free Concentration analysis
Sensor development for testing