Anti-HIV-1 gp120 Recombinant Antibody (C108G) (CAT#: MRO-116LC)

Figure 1 Effects of reduction and deglycosylation on stability of epitopes in the V2 and other domains.

Recombinant BaL gp120 was captured on wells of a 96-well ELISA plate either untreated (open squares) or after deglycosylation with peptide N-glycosidase F (PNGase F; closed diamonds) or reduction by DTT (closed triangles). The reactivity of the indicated MAbs to control or treated rgp120 was titrated for 1 h at 37°C and bound antibody detected with alkaline phosphatase-conjugated anti-IgG, followed by incubation with alkaline phosphatase substrate. OD405, optical density at 405 nm.

The C108g Epitope in the V2 Domain of gp120 Functions as a Potent Neutralization Target When Introduced into Envelope Proteins Derived from Human Immunodeficiency Virus Type 1 Primary Isolates

Figure 2 Effects of mutations in the JR-FL V2 domain on neutralization by V2-specific MAbs C108g and 10/76b and MAbs to the V3 region (4117C) and CD4-binding domain (5145A).

Neutralization assays were performed with env deleted, luc-expressing NL4-3 provirus pseudotypes with either the parental JR-FL Env (closed triangles) or the JR(GKV) variant expressing the C108g epitope in the JR-FL background (open triangles).

The C108g Epitope in the V2 Domain of gp120 Functions as a Potent Neutralization Target When Introduced into Envelope Proteins Derived from Human Immunodeficiency Virus Type 1 Primary Isolates

Figure 3 Analysis of neutralizing activities of MAbs against various domains for the viruses pseudotyped with the two parental Envs SF162 (closed circles) and JR-FL (closed triangles) and with the two C108g-expressing variants SF(NI GKV) (open circles) and JR(GKV) (open triangles).

The MAb used and the domain it recognizes are indicated at the top of each panel.

The C108g Epitope in the V2 Domain of gp120 Functions as a Potent Neutralization Target When Introduced into Envelope Proteins Derived from Human Immunodeficiency Virus Type 1 Primary Isolates

Figure 4 Fluorescence-activated cell sorter assays of the inhibition of binding of either gp120 to U87 cells expressing CD4 by MAbs or sCD4.

Env proteins were used at 1.0 g/ml, MAbs at 20 g/ml, and sCD4 at 100 g/ml. Binding of Env proteins to cells was detected by subsequent staining with biotinylated HIVIG, followed by R-phycoerythrin-conjugated streptavidin and quantitated on a Becton-Dickinson FACScalibur flow cytometer. The MAb used and the extent of inhibition compared to the control in the absence of antibody, measured as the decrease in mean fluorescence intensity compared to the control, are indicated in each panel.

The C108g Epitope in the V2 Domain of gp120 Functions as a Potent Neutralization Target When Introduced into Envelope Proteins Derived from Human Immunodeficiency Virus Type 1 Primary Isolates