Anti-Mouse IgG Alpha Donor beads, 1 mg

The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.

Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.

Technical advancements in antibody engineering has brought about greater interest in more novel antibody therapeutic design and the emergence of new classes of antibody therapeutics called bispecific antibodies (bsAbs). The principle behind bispecific antibody design is to create an antibody / antibody fragment to two or more binding sites to help with the treatment of complex diseases.

As more bsAbs are produced as therapeutics, fast and accurate methods for functionally evaluating and characterizing the stability of these antibodies are necessary during both discovery and development stages, as well as during formulation and quality analysis.

In this application note, we demonstrate how AlphaLISA^® assay technology with the EnVision^® multimode plate reader can be used for bispecific antibody detection, through an example application to characterize the binding and specificity of a mouse bispecific antibody targeting mouse TIGIT and mouse PD-L1.

You will find out:

• What bispecific antibodies are and how they differ from classical antibody formats
• How to design quick and easy experiments to measure binding of the bispecific antibody to the target proteins
• How a no-wash multiplex assay can show direct binding of each site to its respective target simultaneously in one well
• How these assays could be used to evaluate the stability and functional reproducibility between batches of the bsAb preparations

The interactions and bindingof proteins are implicated in a large number of biological processes. The needfor an efficient, highly sensitive assay to study large protein interactions is increasingly important. Alpha Technology is a highly flexible, homogeneous, no-wash assay ideal for the measurement of protein interactions and complexes as large as 200 nm in size

Alpha has been used to study a wide variety of interactions, including protein:protein, protein:peptide, protein:DNA, protein:RNA, protein:carbohydrate, protein:small molecule, receptor:ligand, and nuclear receptor:ligand interactions. Both cell-based and biochemical interactions have been monitored, and applications such as phage display, ELISA, and EMSA (electrophoretic mobility shift assay) have been adapted to Alpha.

This guide presents the simple conversion of an ELISA or other immunoassay to an AlphaLISA® immunoassay.

AlphaScreen® and AlphaLISA® are bead-based assay technologies used to study biomolecular interactions in a microplate format. The acronym “Alpha” stands for Amplified Luminescent Proximity Homogeneous Assay. The assay does not require any washing steps. Binding of proteins or other binding partners captured on the beads leads to an energy transfer from one bead to the other, ultimately producing a luminescent signal.