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Immunodeficiency Virus (HIV) p24 is the 231 amino acid phosphorylated protein of the capsid forming the conical core of the virus that encapsulates the genomic RNA-nucleocapsid complex. p24 is a cleavage product of the p55 Gag polyprotein by viral proteases. HIV p24 and its 55 kDa precursor play a crucial role in the assembly, maturation, and disassembly of HIV. p24 can often be detected two weeks after infection. Subsequently, p24 antibody is produced and complexes with soluble p24 antigen, rendering it undetectable without first dissociating the antibody-antigen complex. Free antigen reappears later in the course of the illness as p24 antibody levels decline. p24 is frequently used for HIV detection in blood, serum samples, and other bodily fluids in acute HIV seroconversion, in neonatal infection, and for monitoring of responses to antiviral drug therapy.
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||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||500 assay points|
The main vectors of gene therapy in research are viruses. The most popular tool for gene delivery is a genetically modified lentivirus. Modified lentivirus (HIV-1) vectors retain their ability to infect undivided cells, thereby increasing their ability to transduce a wide variety of cells, including those that are difficult to transduce. This advantage enables the stable long-term expression of a transgene.
In immunotherapy, CAR-T cells are manufactured by transducing the CAR gene with an HIV-1 vector in T cells to express a specific chimeric p24 protein on their surface. This allows them to recognize cancer cells and destroy them. These CAR-T cells must be generated individually to treat each patient.
This application note demonstrates a comparative quantification of the p24 titer in a lentiviral GFP control sample using Alliance HIV-1 p24 Antigen ELISA and p24 AlphaLISA immunoassay platforms.
Check out the different sections of this application note:
For the detection of three biomarkers in complex sample matrices, the AlphaLISA and Electrochemiluminescent (ECL) assay technologies were shown to have similar: Assay windows (linear dynamic range), Lower and upper detection limit, Intra-and inter-assay precision (lower % CV) The advantages of using AlphaLISA over ECL are: Shorter total assay duration No wash steps No shaking Lower sample volume requirement for equivalent performance Less expensive instrument and plates required
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.