The AlphaLISA® Human IgG4 Pharmacokinetic Detection Kit is designed for the quantitative determination of human IgG4 in monkey serum and other samples, using a homogeneous (no wash steps, no separation steps) assay.
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This kit was developed to measure human IgG4 monoclonal antibodies in rodent and monkey serum (non-human serums) during pharmacokinetic studies. The antibodies were chosen so that the assay shows negligible cross-reactivity with monkey IgG.
Immunoglobulin G (IgG), a major effector molecule of the humoral immune response, accounts for about 75% of the total immunoglobulins in plasma of healthy individuals whereas IgM, IgA, IgD and IgE, each of which has characteristic properties and functions, constitute the remaining 25%. The basic IgG molecule has a four-chain structure, comprising two identical heavy (H) chains and two identical light (L) chains, linked together by inter-chain disulfide bonds. Four IgG subclasses have been identified: IgG1, IgG2, IgG3 and IgG4. Biotherapeutic antibody drugs, usually IgG1 or IgG4 molecules, are becoming increasingly important to treat debilitating diseases such as cancer and autoimmune disorders. Drug levels need to be accurately measured at various stages of drug development, including early antibody discovery, preclinical research in animals, and commercial manufacturing. The present kit permits detection of human IgG1 in different sample matrices, including monkey serum.
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 Class||Antibody|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
The aim of this work was to compare the performance of the AlphaLISA® amplified luminescent proximity homogeneous assay technology platform and an alternative time-resolved fluorescenceresonance energy transfer (TR-FRET) technology assay platform for the detection of five biomarkers.
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
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.
Enzyme-linked Immunosorbent Assay is the most widely Kits adopted method for detection and quantification of cytokines and other biomarkers. This traditional technology offers good,selectivity, sensitivity and assay versatility; however, it has certain disadvantages such as limited dynamic range and low throughput due to the numerous wash steps.