normal guinea pig serum  (Jackson Immuno)

Journal: bioRxiv

Article Title: Effects and mechanisms of monoclonal and polyclonal human antibodies in protection of humanized mice from HIV-1 challenge

doi: 10.1101/2025.11.12.688003

Figure Lengend Snippet: A. Binding of 10-1074 variants and isotype control mAbs to recombinant gp120 in a multiplex binding assay. B. Neutralization potency of 10-1074 variants and isotype control mAbs to REJO.c in TZM-BL assay. C-F. Effector functions of 10-1074 variants and isotype control mAbs in C3 deposition ( C ), viral lysis in the presence of complement ( D ), phagocytosis ( E ), and ADCC defined as FcγRIIIa signaling ( F ). Error bars indicate standard error of the mean across replicates. G . Structural visualization of human IgG Fc (gray) in complex with human FcRn (white) from PDB 7Q15. E430G (blue) and contact residues (orange) are indicated. H. Levels of 10-1074 detected in blood over time. I . Complement-dependent cytotoxicity (CDC) activity against hemolysin-sensitized sheep red blood cells in serum from indicated controls or mouse strain backgrounds with (gray background) and without humanization. Deionized (DI) water and heat inactivated (56°) mouse serum served as positive and negative controls, respectively.

Article Snippet: Complement active guinea pig serum [Fisher Scientific] was diluted 1:100 in GVB ++ (Sigma) and added to the assay plate and incubated for 30 mins at 37°C.

Techniques: Binding Assay, Control, Recombinant, Multiplex Assay, Neutralization, Lysis, Activity Assay

Journal: bioRxiv

Article Title: Effects and mechanisms of monoclonal and polyclonal human antibodies in protection of humanized mice from HIV-1 challenge

doi: 10.1101/2025.11.12.688003

Figure Lengend Snippet: A. Binding of VRC01 IgG subclass variants and IgG1 isotype control mAbs to recombinant gp120. B. Neutralization potency of VRC01 IgG subclass variants and IgG1 isotype control mAbs to REJO.c in TZM-BL assay. C-E. Effector function of VRC01 subclass variants and isotype control mAbs in ( C ) phagocytosis, ( D ) FcγRIIIa activation, and ( E ) viral lysis in the presence of complement assays. Error bars indicate standard deviation. F. Study design. NSG-BLT mice were serially challenged with a cocktail of REJO.c virus and mAb as illustrated. G . Kaplan-Meier curve of infection over time. Statistical significance of VRC01 treatment compared to isotype control defined by log-rank test. H-I . Viral RNA levels over time. Geometric mean and 95% CI of viral RNA over time in each treatment ( H ) group and ( I ) individual animals. Dotted horizontal line indicates limit of detection. Dotted vertical lines indicate virus challenge. Animals per group indicated in inset at left, and those uninfected at the end of the experiment at right.

Article Snippet: Complement active guinea pig serum [Fisher Scientific] was diluted 1:100 in GVB ++ (Sigma) and added to the assay plate and incubated for 30 mins at 37°C.

Techniques: Binding Assay, Control, Recombinant, Neutralization, Activation Assay, Lysis, Standard Deviation, Virus, Infection

Journal: bioRxiv

Article Title: Effects and mechanisms of monoclonal and polyclonal human antibodies in protection of humanized mice from HIV-1 challenge

doi: 10.1101/2025.11.12.688003

Figure Lengend Snippet: A-F . In vitro antibody activities. A. Binding of pooled human serum polyclonal IgG from seronegative donors (IVIG), seropositive donors (HIVIG), from a single seropositive donor (HIV-008), as compared to VRC01 to recombinant gp120 TRO. B. Neutralization potency against RHPA in TZM-BL assay. C-F. Effector function of pAbs in phagocytosis ( C ), ADCC defined by FcγRIIIa activation ( D ), complement deposition ( E ), and complement-dependent viral lysis ( F ) assays. Error bars indicate standard deviation. G. Study design. One batch of S15-CD34 mice was serially challenged intraperitoneally (IP) with a cocktail of RHPA virus and pAb as illustrated. Animals in the IVIG plus VRC01 group were randomized to either IVIG or HIV-008 after the second challenge. H-J . Detection of viral RNA over time by ( H ) treatment group and ( J ) per intervention. Dotted horizontal line indicates limit of detection. Animals per group are indicated in inset. I . Kaplan-Meier curve of infection over time. Results from the third challenge of animals re-randomized from the IVIG plus VRC01 group after the second challenge were included with newly assigned groups at challenge one. Statistical significance of IVIG plus VRC01 and HIV-008 treatment compared to IVIG defined by log-rank test. Censored animals indicated by symbols. Dotted vertical lines indicate times of virus challenge.

Article Snippet: Complement active guinea pig serum [Fisher Scientific] was diluted 1:100 in GVB ++ (Sigma) and added to the assay plate and incubated for 30 mins at 37°C.

Techniques: In Vitro, Binding Assay, Recombinant, Neutralization, Activation Assay, Lysis, Standard Deviation, Virus, Infection