HIV-1 p24 antibody

HIV-1 p24 Mouse antibody
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Cat. No.
BT7508
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

Structure and Function of HIV-1 p24 Antibody

  • Key characteristics:

    • Targets conserved regions of p24, including the CypA binding loop and C-terminal helices .

    • Cross-reactive across HIV-1 subtypes (e.g., subtypes B, C, CRF01_AE) due to conserved epitopes .

Diagnostic Applications

Fourth-generation HIV tests combine p24 antigen and antibody detection, shortening the diagnostic window period to 14–21 days post-exposure .

Performance Metrics of p24 Antibody-Based Assays

Assay TypeDetection LimitSpecificityCross-ReactivitySource
Colloidal Gold Immunoassay (GICA)25 pg/mL98.03%HIV-1 subtypes
Europium Nanoparticle TRF Assay0.5–1 pg/mL>99%All major subtypes
Fourth-Generation ELISA~50 pg/mL99.8–100%Most subtypes

  • Monoclonal antibodies (mAbs):

    • 24-4: Mouse IgG2b mAb used in Western blot, flow cytometry, and immunofluorescence .

    • C65690M/ANT-152: Broadly cross-reactive mAbs for europium nanoparticle assays, detecting 0.5–1,000 pg/mL of p24 .

Immune Response Dynamics

Anti-p24 antibody profiles shift during infection:

  • Acute phase: 86% of patients show polyclonal responses to multiple linear epitopes .

  • Chronic phase: 60% react with single epitopes (e.g., helix 10/11), while 30% lose reactivity .

Dominant Epitopes:

  1. CypA binding loop (helix 4/5) .

  2. C-terminal domain (helix 10/11) .

Therapeutic Potential

Conjugated p24 antibodies show promise in inhibiting viral replication:

  • κFGF-MTS-anti-p24 mAbs: Internalize into T cells, reducing HIV-1 replication by 49–73% in vitro .

  • CAP-1 small molecule inhibitors: Disrupt capsid assembly by targeting p24 .

Challenges and Innovations

  • False negatives: Occurs with divergent HIV strains due to antibody specificity gaps .

  • Novel assays: Dual antibody systems (e.g., C65690M + ANT-152) improve sensitivity for low-p24 samples .

  • Point-of-care tools: GICA enables rapid testing with 98% specificity but requires optimization to reduce false positives .

Research Advancements

  • Nanoparticle-based assays: Europium-labeled antibodies achieve 10–100x higher sensitivity than ELISA .

  • Epitope mapping: Identified immunodominant regions for vaccine design .

Product Specs

Introduction
Human immunodeficiency virus (HIV) is a type of virus known as a retrovirus. It weakens the body's defense system, making individuals vulnerable to infections. HIV attacks specific cells within the immune system, primarily a type of white blood cell known as a helper T cell (specifically CD4+ T cells), as well as macrophages and dendritic cells. The virus reduces the number of CD4+ T cells through three primary ways: directly killing infected cells, increasing the rate of programmed cell death (apoptosis) in infected cells, and enabling the destruction of infected CD4+ T cells by other immune cells called CD8 cytotoxic lymphocytes. When the CD4+ T cell count drops significantly, the body's ability to fight off infections is compromised, making individuals susceptible to a range of illnesses. HIV is classified as a lentivirus, belonging to the Retroviridae family. Lentiviruses share common characteristics in terms of their structure and behavior. They are known to cause long-lasting diseases with extended periods before symptoms appear. These viruses are transmitted as single-stranded, positive-sense RNA viruses enclosed within a protective envelope. Once inside a host cell, the virus's RNA genome is converted into double-stranded DNA by an enzyme called reverse transcriptase, which is carried within the virus particle. This viral DNA is then incorporated into the host cell's DNA using another viral enzyme called integrase, allowing the virus's genetic material to be transcribed. After infecting a cell, the virus can follow two paths: it can either remain dormant (latent) within the cell while the cell continues to function normally, or it can become active and replicate, producing a large quantity of new virus particles that can infect other cells.
Formulation
The antibody is supplied in a liquid solution containing 1 milligram of antibody per milliliter of phosphate-buffered saline (PBS) after it has been reconstituted.
Shipping Conditions
The antibody is shipped in a freeze-dried (lyophilized) form at room temperature.
Storage Procedures
For long-term storage in lyophilized form, keep at 4 degrees Celsius in a dry environment. After reconstitution, if not used within a month, divide into smaller portions and store at -20 degrees Celsius.
Solubility
To reconstitute the antibody, add sterile water (H2O). Mix the solution gently, ensuring the sides of the vial are rinsed, and allow it to sit undisturbed for 30-60 seconds before use.
Titer
When tested using a direct ELISA (enzyme-linked immunosorbent assay) against the recombinant gp24 protein, a 1:10,000 dilution of the antibody produces an optical density (O.D.) reading of 0.4. This measurement is taken using an alkaline phosphatase-conjugated rabbit anti-mouse immunoglobulin G (IgG) secondary antibody from Jackson Laboratories.
Purification Method
Ion exchange column.
Type
Mouse antibody Monoclonal.
Clone
YDHIV1gp24.
Immunogen
r.HIV-1p24.
Ig Subclass
mouse IgG1.

Q&A

FAQs for Researchers on HIV-1 p24 Antibodies in Academic Contexts

Advanced Research Questions

  • How do anti-lipid antibodies (e.g., 11.31, CL1) inhibit HIV-1 infection via p24-independent mechanisms?

    • Mechanistic Insight: Anti-phospholipid antibodies bind monocytes, inducing CCR5-binding chemokines (MIP-1α/MIP-1β), which block R5-tropic HIV-1 entry. Pretreatment of monocytes with 11.31 reduced infection by 87% vs. 35% in CD4+ T-cells .

    • Experimental Validation:

      • Figure 8 ( ): Monocyte pretreatment with 11.31 caused 87% reduction in p24 production post-HIV-1 B.6535 infection.

      • Figure 10 ( ): Neutralizing anti-chemokine antibodies reversed inhibition, confirming chemokine-mediated activity.

  • What strategies resolve contradictions in p24 antibody efficacy across HIV-1 subtypes?

    • Approach:

      • Use antibody pairs targeting conserved epitopes (e.g., C65690M + ANT-152) to mitigate subtype variability .

      • Compare antibody performance in PBMCs from diverse donors to assess strain-specific neutralization (e.g., IgG1b12 inhibited 95% of PBMC infections vs. 4E10 at 40%) .

    • Data Table:

      AntibodyInhibition Rate (%)Mean IC80 (μg/mL)
      IgG1b12959.2
      4E104020.4
      HIVIG98780
      [Source: ]

  • How can p24 antibody assays distinguish acute vs. established HIV-1 infection?

    • Method: Pair p24 antigen detection with antibody screening (e.g., Determine™ HIV-1/2 Ag/Ab Combo). Acute infection is indicated by p24+/Ab– results, while established infection shows p24+/Ab+ .

    • Technical Note: Use streptavidin-based capture in the Upper Test Area to avoid cross-reactivity with HIV-2 p26 .

Methodological Challenges and Solutions

  • Why do some p24 antibodies fail to detect emerging HIV-1 strains?

    • Root Cause: Epitope variability in the capsid protein (e.g., subtype-specific polymorphisms in p24).

    • Solution: Screen hybridoma libraries from autoimmune patients (e.g., APAS, lupus) for cross-reactive anti-lipid antibodies .

  • How can p24 assays be adapted for resource-limited settings?

    • Innovation: Replace RNA quantification with boosted p24 antigen assays (cost: ~1/10th of RT-PCR). For infant diagnosis, use heat-denatured plasma to bypass RNA testing .

Data Interpretation Guidelines

  • False Positives: Exclude samples from patients on biotin supplements .

  • Quantification: Linearize p24 signals using log-transformed TRF values for low-abundance antigens .

  • Cross-Validation: Compare p24 levels with RNA viral loads (≥0.67 correlation coefficient required) .

Product Science Overview

Introduction

HIV-1 p24 is a core protein of the Human Immunodeficiency Virus type 1 (HIV-1), which is a major causative agent of Acquired Immunodeficiency Syndrome (AIDS). The p24 protein is a part of the Gag polyprotein and plays a crucial role in the assembly and maturation of the virus. Mouse antibodies against HIV-1 p24 are widely used in research and diagnostic applications due to their specificity and effectiveness in detecting the p24 antigen.

Structure and Function of HIV-1 p24

The p24 protein is a capsid protein that forms the conical core of the HIV-1 virion. It is composed of approximately 231 amino acids and has a molecular weight of around 24 kDa. The p24 protein is essential for the formation of the viral capsid, which encases the viral RNA and enzymes necessary for the replication of the virus. The protein is highly conserved among different HIV-1 strains, making it an ideal target for diagnostic assays .

Importance in HIV Diagnosis

The detection of p24 antigen is a critical component in the early diagnosis of HIV infection. The p24 antigen can be detected in the blood of infected individuals before the appearance of antibodies, making it a valuable marker for early HIV detection. The p24 antigen test is also used to monitor the viral load in patients undergoing antiretroviral therapy, as the levels of p24 correlate with the amount of virus present in the body .

Mouse Antibodies Against HIV-1 p24

Mouse antibodies against HIV-1 p24 are monoclonal antibodies produced by immunizing mice with the p24 protein. These antibodies are highly specific to the p24 antigen and are used in various immunoassays, including Western Blot, ELISA, Immunohistochemistry, and Immunofluorescence . The use of mouse antibodies allows for the precise detection and quantification of the p24 protein in different biological samples.

Applications in Research and Diagnostics
  1. Western Blot: Mouse antibodies against HIV-1 p24 are used in Western Blot assays to detect the presence of p24 protein in lysates of infected cells. This technique is commonly used in research to study the expression and processing of the Gag polyprotein.
  2. ELISA: Enzyme-Linked Immunosorbent Assay (ELISA) is a widely used technique for the quantitative detection of p24 antigen in serum or plasma samples. Mouse antibodies are used as capture and detection antibodies in these assays.
  3. Immunohistochemistry: This technique involves the use of mouse antibodies to detect p24 protein in tissue sections. It is used to study the distribution and localization of the virus in infected tissues.
  4. Immunofluorescence: Mouse antibodies conjugated with fluorescent dyes are used to visualize the p24 protein in infected cells under a fluorescence microscope. This technique is useful for studying the intracellular localization and trafficking of the virus .

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