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AlphaPlex®-645 (Samarium) Acceptor beads conjugated to anti-6XHis antibody. The mouse monoclonal antibody detects proteins containing accessible consecutive histidine residues at the amino or carboxy terminus. These beads can be used to capture His-tagged proteins and peptides, and can be used with Alpha Donor beads to create AlphaPlex no-wash assays for:
AlphaPlex Acceptor beads are intended to be used in multiplexing assays, in conjunction with AlphaLISA Acceptor beads and Alpha Donor beads. In a typical Alpha assay, 250 μg of Acceptor beads is sufficient to run 250-500 wells using a 50 μL reaction volume.
|Antibody Conjugates||Anti-6X His|
|Bead Type or Core Bead Type||AlphaPlex Samarium (645) Acceptor|
|Experimental Type||In vitro|
|Product Brand Name||AlphaPlex|
|Shipping Condition||Blue Ice|
|Unit Size||250 μg|
Anti-inflammatory monoclonal antibody drugs that specifically target TNFα, such as Humira®, have been highly successful in the market. As patents expire on these top-selling drugs, effort has been placed on developing biosimilars. Biosimilars differ from small molecule generic drugs in that their chemical structure does not have to be exactly the same as the patented drug. Therefore, the FDA has stringent requirements for proving that the biosimilars have the same efficacy and safety profile as the patented drug. Companies that develop biosimilars are tasked with proving that the biosimilar shows equivalent pharmacokinetics as the patented drug.
Proving “biosimilarity” involves comparing parameters such as overall exposure, absorption, half-life, and clearance time using patient samples. Sensitive, robust, and fast assays are needed to measure these parameters. Traditional methods for detecting and quantifying these drugs in patient samples include time-consuming, wash-based ELISA and MSD methods. In contrast, AlphaLISA allows for fast, no-wash, high-throughput detection and quantification of the drug of interest in a variety of sample matrices. Here, we demonstrate the application of AlphaLISA for detecting biosimilars targeting TNFα.
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.