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Protein A is a 42 kDa protein widely used in affinity chromatography to purify antibodies, most importantly antibody therapeutics. The native form of Protein A from S. Aureus, in addition to many modified recombinant Protein A variants, are used in chromatography. The strong affinity and the high selectivity of Protein A for antibodies lead to an effective removal of host cell proteins. However, Protein A can be leached from columns to contaminate the antibody preparation. Since Protein A could elicit mitogenic or immunological reactions, there are strict regulations as to the acceptable levels of Protein A in antibody preparations. Therefore, there is a current need to quantitate Protein A in antibody preparations with excellent sensitivity and robustness, while maintaining high throughput capabilities.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
|Assay Target||Residual Protein A|
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||500 assay points|
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.