| Description | This mouse anti-HIV-1 gp120 neutralizing antibody (clone 102) recognizes the conserved CD4 binding region of HIV-1 gp120. It has been tested for use in ELISA, IB and IF. Antibody 102 can neutralize the HIV-1 in infected human MT4 T-lymphoid cells. Antibody 102 has inhibition of replication of various strains of HIV-1, including primary isolates of HIV-1(SF2) B subtypes. |
| Clonality | Monoclonal |
| Host Species | Mouse |
| Target Species | HIV-1 |
| Epitope | The conserved CD4-binding region of gp120 |
| Neutralization Mechanism | Interference with viral replication and inhibition of viral infection. |
| IC50 | 20 nM for HIV-1 SF2 |
| IC100 | 40 µg/ml |
| Affinity | IC50 = 20 nM (mAb); IC50 = 160 nM (scFv) |
| Function | Envelope glycoprotein gp160: Oligomerizes in the host endoplasmic reticulum into predominantly trimers. In a second time, gp160 transits in the host Golgi, where glycosylation is completed. The precursor is then proteolytically cleaved in the trans-Golgi and thereby activated by cellular furin or furin-like proteases to produce gp120 and gp41. Surface protein gp120: Attaches the virus to the host lymphoid cell by binding to the primary receptor CD4. This interaction induces a structural rearrangement creating a high affinity binding site for a chemokine coreceptor like CXCR4 and/or CCR5. Acts as a ligand for CD209/DC-SIGN and CLEC4M/DC-SIGNR, which are respectively found on dendritic cells (DCs), and on endothelial cells of liver sinusoids and lymph node sinuses. These interactions allow capture of viral particles at mucosal surfaces by these cells and subsequent transmission to permissive cells. HIV subverts the migration properties of dendritic cells to gain access to CD4+ T-cells in lymph nodes. Virus transmission to permissive T-cells occurs either in trans (without DCs infection, through viral capture and transmission), or in cis (following DCs productive infection, through the usual CD4-gp120 interaction), thereby inducing a robust infection. In trans infection, bound virions remain infectious over days and it is proposed that they are not degraded, but protected in non-lysosomal acidic organelles within the DCs close to the cell membrane thus contributing to the viral infectious potential during DCs' migration from the periphery to the lymphoid tissues. On arrival at lymphoid tissues, intact virions recycle back to DCs' cell surface allowing virus transmission to CD4+ T-cells. Transmembrane protein gp41: Acts as a class I viral fusion protein. Under the current model, the protein has at least 3 conformational states: pre-fusion native state, pre-hairpin intermediate state, and post-fusion hairpin state. During fusion of viral and target intracellular membranes, the coiled coil regions (heptad repeats) assume a trimer-of-hairpins structure, positioning the fusion peptide in close proximity to the C-terminal region of the ectodomain. The formation of this structure appears to drive apposition and subsequent fusion of viral and target cell membranes. Complete fusion occurs in host cell endosomes and is dynamin-dependent, however some lipid transfer might occur at the plasma membrane. The virus undergoes clathrin-dependent internalization long before endosomal fusion, thus minimizing the surface exposure of conserved viral epitopes during fusion and reducing the efficacy of inhibitors targeting these epitopes. Membranes fusion leads to delivery of the nucleocapsid into the cytoplasm. |
| Isotype | Mouse IgG2a |
| Expression Species | HEK293F or CHO cell line |
| Conjugation | Unconjugated |
| Purity | >95% |
| Endotoxin | <1 EU/mg |
| Form | Liquid |
| Purification | Protein G purified |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4°C within a week. For longer storage, aliquot and store at -20°C. |
| Application | ELISA; IB; IF; Neut |
| Application Notes | The binding specificity of this antibody to HIV-1 antigens was analyzed by ELISA, Immunoblot and Immunofluorescence assay. The potency of HIV-1 neutralization by mAb102 was assessed in infected human MT4 T-lymphoid cells. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
| Post Translational Modification | Palmitoylation of the transmembrane protein and of Env polyprotein (prior to its proteolytic cleavage) is essential for their association with host cell membrane lipid rafts. Palmitoylation is therefore required for envelope trafficking to classical lipid rafts, but not for viral replication. Highly glycosylated by host. The high number of glycan on the protein is reffered to as 'glycan shield' because it contributes to hide protein sequence from adaptive immune system. Specific enzymatic cleavages in vivo yield mature proteins. Envelope glycoproteins are synthesized as an inactive precursor that is heavily N-glycosylated and processed likely by host cell furin in the Golgi to yield the mature SU and TM proteins. The cleavage site between SU and TM requires the minimal sequence [KR]-X-[KR]-R. About 2 of the 9 disulfide bonds of gp41 are reduced by P4HB/PDI, following binding to CD4 receptor. |
