• Detection technology : Surface Plasmon Resonance (SPR) biosensor
• Information provided : Kinetic and affinity data (K_D , k_a, and k_d ), 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 : 40µm
• Sample/reagent capacity : 1 x 96-or 384-well microplate and up to 33 reagent vials
• Analysis temperature : 4^oC to 45^oC (maximum 20^oC below range ambient temperature)
• Sample storage : 4^oC to 45^oC (maximum 15^oC below ambient temperature)
• Sample refractive index range : 1.33 to 1.40
• Buffer selector : Automatic switching between 4 buffers
• In-line reference subtraction : Automatic

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^oC to 45^oC
• 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

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.




The system can be used to study interactions involving any kind of molecule, from organic drug candidates to proteins, nucleic acids, glycoproteins and even viruses and whole cells. Since the response is a measure of the change in mass concentration, the response per molar unit of interactant is proportional to the molecular weight (smaller molecules give lower molar responses). The detection principle does not require any of the interactants to be labeled, and measurements can be performed on complex mixtures such as cell culture supernatants or cell extracts as well as purified interactants. The identity of the interactant monitored in a complex sample matrix is determined by the interaction specificity of the partner attached to the surface. The SPR detection principle is non-invasive and works equally well on clear and colored or opaque samples. The system uses a range of sensor surfaces with a controlled flow system (comprising 4 flow cells) for delivery of samples and reagents to the sensor surface. 

Publications:

• Rane, J. S., Kumari, A., and Panda, D. (2019). An acetylation mimicking mutation, K274Q, in tau imparts neurotoxicity by enhancing tau aggregation and inhibiting tubulin polymerization. Biochemical Journal, 476(10), 1401-1417. doi: 10.1042/BCJ20190042

• Sharma K, Mehra S, Singh Sawner A, Markam PS, Panigrahi R, Navalkar A, Chatterjee D, Kumar R, Kadu P, Patel K, Ray S, Kumar A and Maji SK (2020), Effect of disease-associated P123H and V70M mutations on β-synuclein fibrillation. ACS Chem Neuroscience, 16; 11(18):2836-2848. DOI: 10.1021/acschemneuro.0c00405.

• Harish M, Venkatraman P. Evolution of biophysical tools for quantitative protein interactions and drug discovery Emerg Top Life Sci. 2021 May 14;5(1):1-12. doi: 10.1042/ETLS20200258.PMID: 33739398

• Chatterjee1 , R.S. Jacob1 , S. Ray1 , A. Navalkar1 , N. Singh1 , S. Sengupta1,2 , L. Gadhe1 4 , P. Kadu1 , D. Datta1 , A. Paul1 , A. Sakunthala1 , S. Mehra1 , C. Pindi3 , S. Kumar4 , P. S. Singru4 5 , S. Senapati3 and S. K. Maji (2022),Co-aggregation and secondary nucleation in the life cycle of human prolactin/galanin 2 functional amyloids 3

• The paper entitled 'Mechanistic physicochemical insights into glycation and drug transport by serum albumin: Implications in diabetic conditions' is not published yet. It is recommended for revision in Biochimie

Conference:

• Nag A., Pittu Sandhya Rani and Mehra, S., “SCO4122 is a redox responsive regulator of Mar R family that regulates the efflux pump SCO4121 in Streptomyces coelicolor", ASM Microbe, June 20- June 24, 2021, Online

Publications:

• Rane, J. S., Kumari, A., and Panda, D. (2019). An acetylation mimicking mutation, K274Q, in tau imparts neurotoxicity by enhancing tau aggregation and inhibiting tubulin polymerization. Biochemical Journal, 476(10), 1401-1417. doi: 10.1042/BCJ20190042

• Sharma K, Mehra S, Singh Sawner A, Markam PS, Panigrahi R, Navalkar A, Chatterjee D, Kumar R, Kadu P, Patel K, Ray S, Kumar A and Maji SK (2020), Effect of disease-associated P123H and V70M mutations on β-synuclein fibrillation. ACS Chem Neuroscience, 16; 11(18):2836-2848. DOI: 10.1021/acschemneuro.0c00405.

• Harish M, Venkatraman P. Evolution of biophysical tools for quantitative protein interactions and drug discovery Emerg Top Life Sci. 2021 May 14;5(1):1-12. doi: 10.1042/ETLS20200258.PMID: 33739398

• Chatterjee1 , R.S. Jacob1 , S. Ray1 , A. Navalkar1 , N. Singh1 , S. Sengupta1,2 , L. Gadhe1 4 , P. Kadu1 , D. Datta1 , A. Paul1 , A. Sakunthala1 , S. Mehra1 , C. Pindi3 , S. Kumar4 , P. S. Singru4 5 , S. Senapati3 and S. K. Maji (2022),Co-aggregation and secondary nucleation in the life cycle of human prolactin/galanin 2 functional amyloids 3

• The paper entitled 'Mechanistic physicochemical insights into glycation and drug transport by serum albumin: Implications in diabetic conditions' is not published yet. It is recommended for revision in Biochimie

Conference:

• Nag A., Pittu Sandhya Rani and Mehra, S., “SCO4122 is a redox responsive regulator of Mar R family that regulates the efflux pump SCO4121 in Streptomyces coelicolor", ASM Microbe, June 20- June 24, 2021, Online

• 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 que 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.

• Only online sample registration through the IRCC webpage will be accepted.
• The user will be informed first about the meeting date and time, and later about the experiment date via email.
• 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.