Currently, therapeutic monoclonal antibodies (mAbs) are more and more popular in treating diseases such as cancer. Good therapeutic antibody candidates should have high affinity and specificity for the target. There is a direct correlation between the target binding affinity and the efficacy of the therapeutic antibody. High-affinity means better targeting ability and enhanced antibody-dependent effects. Several methodologies have been established in recent years for the determination of the target binding affinity of antibodies. Creative Biolabs offers various technologies for target binding affinity measurement, such as Surface Plasmon Resonance (SPR), ELISA-based Assay, and Flow Cytometry Assay.
SPR is a rapidly developing technique for the measurement of the kinetics and binding affinities of ligand binding interactions. SPR detects the resonant oscillation of conduction electrons in the immediate vicinity of the surface layer of a sensor chip in a real-time monitoring and label-free manner. These oscillations are very sensitive to any change of this boundary of the conductor and the external medium. Therefore, binding of molecules to the conducting surface will lead to changes of resonant oscillation and can be detected by the SPR technology.
Figure 1. The schematic diagram of Surface Plasmon Resonance (SPR).
The enzyme-linked immunosorbent assay (ELISA) is a widely used solid-phase enzyme immunoassay (EIA) that detects and quantifies substances (e.g., antigen proteins or antibodies) based on color change. ELISA-based assay is a useful tool for determining serum antibody concentrations and antigen-antibody interactions. By detecting the concentration of bound antibodies in an ELISA-based assay, the binding affinity of the antibody to the target antigen (known concentration) can be verified.
Figure 2. Schematic representation of the paper-based ELISA assay developed for the diagnosis of canine leishmaniasis (Costa, M. N., 2014).
Flow cytometry is a biophysical technology that has been employed in many fields, including biomarker/bio-label detection, cell counting, cell sorting, and etc. In this technology, cells are suspended in a stream of fluid, pass through an electronic detection apparatus, and are detected in a laser- or impedance-based method. Flow cytometry assays allow massive analysis of the physical and chemical characters of tons of particles per second. Cell-based flow cytometry assay can also be utilized for antibody affinity measurement. One advantage of this approach is that it allows antigen folding and antigen-antibody interaction in a natural environment. Another significant advantage is that antibodies used in this assay do not require the purification procedure.
Target binding affinity is crucial for the in vivo efficacy of therapeutic antibodies. With our well-established BsAbs analysis platform, the experienced scientists at Creative Biolabs are dedicated to helping you develop BsAbs. Our high quality target binding affinity measurement services will greatly contribute to the successes of your projects. Creative Biolabs also provides various other services regarding BsAbs development. Please feel free to contact us for more information and a detailed quote.
References
1. Willets, K. A.; Van Duyne R. P. Localized surface plasmon resonance spectroscopy and sensing. Annu. Rev. Phys. Chem. 2007, 58: 267-297.
2. Mendoza, L. G.; et al. High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA). Biotechniques. 1999, 27: 778-779.
3. Costa, M. N.; et al. A low cost, safe, disposable, rapid and self-sustainable paper-based platform for diagnostic testing: lab-on-paper. Nanotechnology. 2014, 25(9): 094006.
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