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 >  Protein>HLA-A*0201 | B2M & HIV Gag >HLH-H82E8

Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (Monomer, MALS verified)

更有Streptavidin Protein-PESTN-NP117STN-NP119)快速支持您想要的MHC Tetramer组装!

表达区间及表达系统(Source)

Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (HLH-H82E8) is expressed from human 293 cells (HEK293). It contains AA Gly 25 - Ile 308 (HLA-A*02:01) & Ile 21 - Met 119 (B2M) & SLYNTVATL peptide (Accession # AAA59606.1 (HLA-A*02:01) & P61769 (B2M) & SLYNTVATL).

Predicted N-terminus: Gly 25 & Ser

Request for sequence

蛋白结构(Molecular Characterization)

Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein is produced by co-expression of HLA and B2M loaded with HIV Gag peptide.

This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).

The protein has a calculated MW of 36.3 kDa and 13.7 kDa. The protein migrates as 40-43 kDa and 14 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

标记(Labeling)

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

蛋白标记度(Protein Ratio)

Passed as determined by the HABA assay / binding ELISA.

内毒素(Endotoxin)

Less than 1.0 EU per μg by the LAL method.

纯度(Purity)

>95% as determined by SDS-PAGE.

>95% as determined by SEC-MALS.

制剂(Formulation)

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

Contact us for customized product form or formulation.

重构方法(Reconstitution)

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

存储(Storage)

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

This product is stable after storage at:

  1. -20°C to -70°C for 12 months in lyophilized state;
  2. -70°C for 3 months under sterile conditions after reconstitution.

质量管理控制体系(QMS)

  1. 质量管理体系(ISO, GMP)
  2. 质量优势
  3. 质控流程
 

电泳(SDS-PAGE)

HLA-A*0201 | B2M & HIV Gag SDS-PAGE

Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.

SEC-MALS

HLA-A*0201 | B2M & HIV Gag SEC-MALS

The purity of Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (Cat. No. HLH-H82E8) is more than 95% and the molecular weight of this protein is around 48-58 kDa verified by SEC-MALS.

Report

 

活性(Bioactivity)-ELISA

HLA-A*0201 | B2M & HIV Gag ELISA

Immobilized Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (Cat. No. HLH-H82E8) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-HLA class I Antibody, Human IgG1 (W6/32) with a linear range of 0.1-2 ng/mL (QC tested).

Protocol

HLA-A*0201 | B2M & HIV Gag ELISA

Immobilized Biotinylated Human HLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (Cat. No. HLH-H82E8) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

Protocol

 

活性(Bioactivity)-SPR

HLA-A*0201 | B2M & HIV Gag SPR

HIV Gag TCR captured on Protein A Chip can bind Biotinylated HumanHLA-A*02:01&B2M&HIV Gag (SLYNTVATL) Complex Protein (Cat. No. HLH-H82E8) with an affinity constant of 0.345 nM as determined in SPR assay (Biacore 8K) (Routinely tested).

Protocol

 
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背景(Background)

HIV (human immunodeficiency virus) is a virus that attacks the body’s immune system. If HIV is not treated, it can lead to AIDS (acquired immunodeficiency syndrome). It can be transmitted via the exchange of a variety of body fluids from infected people, such as blood, breast milk, semen and vaginal secretions, And from a mother to her child during pregnancy and delivery. This disease can be managed by treatment regimens composed of a combination of antiretroviral (ARV) drugs. The Human HLA-A*0201 HIV (SLYNTVATL) complex protein is a complex of HLA-A*0201 of the MHC Class I, B2M, and SLYNTVATL peptide of the HIV.

 

前沿进展

Therapeutic immunization with human immunodeficiency virus type 1 (HIV-1) peptide-loaded dendritic cells is safe and induces immunogenicity in HIV-1-infected individuals
Connolly, Whiteside, Wilson et al
Clin Vaccine Immunol (2008) 15 (2), 284-92
Abstract: Treatments for human immunodeficiency virus type 1 (HIV-1)-positive individuals that augment HIV-1 suppression and have potential for achieving long-term control of HIV-1 viremia in the absence of antiretroviral therapy (ART) are urgently needed. We therefore conducted a phase I, clinical safety trial of a dendritic cell (DC)-based vaccination strategy as immunotherapy for HIV-1-positive individuals on ART. We studied 18 HIV-1-positive subjects on ART who underwent leukapheresis to obtain peripheral blood mononuclear cells for DC generation from monocytes cultured with cytokines. Mature DC were pulsed with three HIV-1 HLA*A0201 Gag, Env, and Pol peptides and one influenza A virus matrix protein peptide. The vaccine was administered to donors randomized to receive two vaccinations, either intravenously or subcutaneously. The primary end points were safety and tolerability of two doses of peptide-DC vaccine (3 million versus 10 million). Secondary end points included gamma interferon (IFN-gamma) enzyme-linked immunospot assay responses and clinical correlates of an immune response to vaccination. Autologous DC-peptide vaccine was safe, well tolerated, and feasible for use in all participants. Adverse events were rare. Although the trial was not powered to assess an immunologic response, a significantly increased frequency of HIV-1 peptide-specific IFN-gamma-positive cells was observed 2 weeks following the second vaccine, with three individuals responding to all four peptides. DC vaccination was safe, was feasible, and showed promise of immunogenicity in ART-treated, HIV-1-positive individuals. Additional studies of DC immunization strategies for HIV-1 infection are warranted.
Induction of primary anti-HIV CD4 and CD8 T cell responses by dendritic cells transduced with self-inactivating lentiviral vectors
Chen, Wang, Chang
Cell Immunol (2006) 243 (1), 10-8
Abstract: In this study, we demonstrate that a minimal self-inactivating (SIN) lentiviral vector (LV) that does not encode any human immunodeficiency virus (HIV) genes is able to induce HIV-specific CD4 and CD8 T cell responses after transduction of dendritic cells (DCs). The LV-DC-primed T cells displayed HIV-specific lytic degranulation, as illustrated by acquisition of CD107a/b expression on the cell surface and up-regulation of active caspase 3. HIV-specific cytotoxic T lymphocyte (CTL) response was consistently detected using different assays, and T cell receptors specific to three prominent HIV epitopes, SL9 (Gag peptide: SLYNTVATL), IV9 (Pol peptide: ILKEPVHGV), and MA10 (In peptide: MASDFNLPPV) were detected using HLA-A0201 peptide-tetramers. These results demonstrate that DCs transduced with the minimal SIN-LV can efficiently induce HIV-specific CD4 and CD8 T cell responses. Since LVs are popular gene transfer tools, our results have fundamental implications for future LV applications and DC vaccine development.
Phenotype and functional characteristics of HIV-specific cytotoxic CD8+ T cells in chronically infected patients: dual effects of highly active antiretroviral therapy
Benito, López, Lozano et al
J Acquir Immune Defic Syndr (2003) 34 (3), 255-66
Abstract: Cytotoxic CD8+ T cells (CTLs) are critical for the control of viral infections. Although these cells can be recognized in most HIV-infected individuals, they fail to successfully control HIV replication. Distinct functional defects seem to limit their efficacy in HIV infection, although they have been not fully elucidated.Blood lymphocytes collected from 61 HLA-A0201+, untreated, chronically HIV-infected individuals were examined for the presence of CTLs against epitopes from HIV Gag and Pol proteins, using tetrameric complexes. Several functional aspects of these cells were further analyzed (immunophenotype; ability to produce interferon-gamma (IFN-gamma) in response to the specific peptide; proliferative capacity; and cytolytic activity). Lymphoproliferative responses of these cells confronting different stimulus were also evaluated. A longitudinal analysis was carried out in a subgroup who underwent antiretroviral therapy and were followed for 6 months.CD8+ T cells staining with the tetramer complexes (Tet+) were detected in 44% of patients, with TetGag+ cells being more frequently detected and at higher levels than TetPol+ cells. Most Tet+ cells expressed a memory phenotype, showed an impaired ability to produce IFN-gamma when stimulated with the cognate peptide, and showed a very low expansion when cultured in the presence of the peptide. There was a negative correlation between the proportion of Tet+ cells producing IFN-gamma and plasma HIV-RNA. Although Tet+ cells diminished in most individuals after beginning antiretroviral therapy, some patients showed de novo appearance of Tet+ cells.Most Tet+ cells in chronic HIV-infected individuals express a memory phenotype and show an impaired production of IFN-gamma and a lower proliferative response to specific HIV antigens. Interestingly, some individuals under successful antiretroviral therapy may show de novo appearance of specific CTLs. The implications of these findings are relevant for a better understanding of virus-host interactions.
Distinct molecular mechanisms account for the specificity of two different T-cell receptors
Anikeeva, Lebedeva, Krogsgaard et al
Biochemistry (2003) 42 (16), 4709-16
Abstract: Analysis of the thermodynamics of the interactions between the D3 T-cell receptor (TCR) and its natural ligand, an HIV peptide bound to a HLA-A0201 (HLA-A2) major histocompatibility complex (MHC) protein, shows both similarities and striking differences when compared with the 2B4 TCR binding to its peptide-MHC ligand. The equilibrium thermodynamic parameters of both reactions are consistent with a conformational adjustment at the binding interface during the formation of specific TCR-peptide-MHC complexes. However, osmolytic reagents that dehydrate protein surfaces have profoundly different effects on the strength of the two reactions, indicating that water molecules make very different contributions-enhancing the binding of D3 TCR but weakening the binding of 2B4 TCR. The use of these different mechanisms by TCRs to recognize ligands might be an important means augmenting their inherent cross-reactivity.
Showing 1-4 of 5 papers.
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