There are a number of technologies that may be utilized to assess antibody antigen binding, i.e. Enzyme-linked immunosorbent assay (ELISA), quartz crystal microbalance (QCM), and Isothermal Titration Calorimetry (ITC). However, few encompass as many advantages as Surface Plasmon Resonance (SPR). SPR enables (1) high sensitivity, (2) label-free detection, (3) real-time monitoring, (4) low volume sample consumption, (5) quantitative evaluation, and (6) determination of kinetic rate constants. Furthermore, SPR is easy to perform and can be a cost-effective solution.
Abzyme custom SPR-based antibody service can help you:
• Rank your monoclonal antibodies based on association/disassociation rates and binding affinity
• Obtain detailed binding kinetic data
• Identify monoclonal antibodies binding to different epitopes of the antigen
Antibody antigen binding affinity will be characterized using fully automated BI-4500 5-Channel SPR System (Biosensing Instrument) that delivers high sensitivity and fast response times with superior flexibility. The BI-4500 system is equipped with BI-DirectFlow™ technology, which enables precision sample delivery with near-zero dispersion for higher-quality results, ideal for the study of ultra-fast kinetic interactions such as those in DNA sequencing, protein-protein interactions, ligand-receptor recognition, and drug development applications. The BI-4500 produces superior, high-quality data at a completely affordable price.
Sensorgram from Biosensing Instrument Application Note.
Key advantages of fully automated BI-4500 5-Channel SPR System are:
• 5-channel SPR detection module for background and reference subtraction, and various antigen concentrations
• Wide dynamic range (greater than 10-5) for both large and small molecules (greater than 100 Daltons) enables users to measure binding constants down to a few pM
• Sample delivery is fully-automated with BI-Autosampler to avoid human errors
• Precision temperature control with broad range (6 to 50oC) for thermodyamic studies
• Broad response time for slow (hours) and fast (greater than 4 ms) kinetic processes
• The BI-4500 system comes complete with a precision syringe pump, PC station, and powerful control/analysis software.
Materials needed for SPR service:
• 50 µg of each sample
• Molecular weights of samples
• Concentration of samples
Special considerations may be required for affinity determination of antigens with repeated or multiple epitopes. Please contact us for more information and quote.
Our cytoflow service allows determination of the interaction of an antibody with different cells to characterize cell specificity and potential target cells before in vivo studies.
Materials needed for cytoflow service:
• 50 µg of each sample
• Specific target cells, although Abzyme has several cells on hand and the availability will be discussed at the initiation of the project
SPR: At the end of the SPR project, our team provides a sensorgram of antibody antigen interaction and a summary of the kinetic data (ka, kd, kD).
Cytoflow analysis: At the end of the cytoflow project, our team provides an analysis of each antibody target cell interaction.
Typically, a SPR project will be completed within two weeks after receiving your materials. Our cytoflow analysis timeline is dependent on the growth rate of your target cells.