mouse anti hbsag monoclonal antibody  (Bio-Rad)

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 1. Optimization of in-house quantitative sandwich ELISA for detection of HBsAg and HBeAg. Standard curve depicting the optimal dilution of detection antibody for the detection of (A) HBsAg and (B) HBeAg. Primary antibodies used for antigen capture were mouse monoclonal α-HBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M). Detection was achieved using HRP-conjugated α-HBsAg goat polyclonal antibody (Fitzgerald, 70-HG15S) and α-HBeAg mouse monoclonal antibody (Fitzgerald, 61-H10K). OD450 values were normalized against BSA control wells. The assay was optimized by adjusting antibody concentrations to achieve a strong signal-to-noise ratio while maintaining linearity. Data were normalized by subtracting the optical density reading at 450 nanometers (OD450nm) from bovine serum albumin (BSA) control wells. (C) Summary table of ELISA data for HBeAg and HBsAg. Assay quality was assessed based on linearity of the standard curve R2 and Z′ factor. Data from 3 biological replicates.

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal αHBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Sandwich ELISA, Control, Enzyme-linked Immunosorbent Assay, HBsAg Assay

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 2. Comparison of quantitative in-house ELISA assays for HBsAg (A) and HBeAg (B) detection in plasma samples from HBV-infected individuals (genotypes A–F), evaluated against a commercial ELISA (Abbott Architect). Dilution series (10-, 100-, 500-, 1000-fold) were conducted to determine the minimum plasma volume required for reliable quantification of HBsAg and HBeAg. Statistical analysis via one-way ANOVA with Tukey’s HSD post hoc test revealed significant differences (**) at 1000-fold dilution between groups (p < 0.05), and no significant (ns) differences between other dilution series. Data are presented as mean ± SD from three technical replicates per sample. (C) Summary of 35 samples tested, detailing HBsAg and HBeAg positivity rates across genotypes (A–F), as determined by the Abbott Architect assay. A 10-fold plasma dilution (10 µL in 100 µL of blocking buffer) reliably quantified (D) HBsAg and (E) HBeAg. Notably, HBeAg levels in samples tested from genotypes A and E were below the lower limit of detection (

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal αHBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Infection, Blocking Assay

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 4. Comparison of various DNA isolation methods for determining the EC50 of TDF in HepAD38 cells. HepAD38 cells were seeded in 24-well plates or 96-well plates and treated with TDF for 14 days with media and drug replenishment every 72 h. (A) To determine the toxicity of TDF, HepAD38 cells in the 96-well plate format were incubated with Alamar Blue for 2 h prior to reading the absorbance at 560 nm and 600 nm. Data were normalized according to the manufacturer’s directions to determine cell viability. (B) HBeAg and HBsAg could be quantified from the supernatant of 96-well HepAD38 cells treated with Alamar Blue. Extracellular (cell supernatant, top panel) and intracellular (bottom panel) HBV DNA were determined in (C) 96-well and (D) 24-well plate formats and plotted as % HBV inhibition against log [TDF] to determine the EC50. To investigate if Alamar Blue negatively impacts EC50 determination, HepAD38 cells from the 96-well plate treated with Alamar Blue were also subjected to DNA isolation and quantification. (E) Summary of TDF EC50 values obtained from the treatment of HepAD38 cells in either a 24- or 96-well format. Assay Z′

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal αHBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Comparison, DNA Extraction, Incubation, Inhibition

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 1. Optimization of in-house quantitative sandwich ELISA for detection of HBsAg and HBeAg. Standard curve depicting the optimal dilution of detection antibody for the detection of (A) HBsAg and (B) HBeAg. Primary antibodies used for antigen capture were mouse monoclonal α-HBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M). Detection was achieved using HRP-conjugated α-HBsAg goat polyclonal antibody (Fitzgerald, 70-HG15S) and α-HBeAg mouse monoclonal antibody (Fitzgerald, 61-H10K). OD450 values were normalized against BSA control wells. The assay was optimized by adjusting antibody concentrations to achieve a strong signal-to-noise ratio while maintaining linearity. Data were normalized by subtracting the optical density reading at 450 nanometers (OD450nm) from bovine serum albumin (BSA) control wells. (C) Summary table of ELISA data for HBeAg and HBsAg. Assay quality was assessed based on linearity of the standard curve R2 and Z′ factor. Data from 3 biological replicates.

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal α-HBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Sandwich ELISA, Control, Enzyme-linked Immunosorbent Assay, HBsAg Assay

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 2. Comparison of quantitative in-house ELISA assays for HBsAg (A) and HBeAg (B) detection in plasma samples from HBV-infected individuals (genotypes A–F), evaluated against a commercial ELISA (Abbott Architect). Dilution series (10-, 100-, 500-, 1000-fold) were conducted to determine the minimum plasma volume required for reliable quantification of HBsAg and HBeAg. Statistical analysis via one-way ANOVA with Tukey’s HSD post hoc test revealed significant differences (**) at 1000-fold dilution between groups (p < 0.05), and no significant (ns) differences between other dilution series. Data are presented as mean ± SD from three technical replicates per sample. (C) Summary of 35 samples tested, detailing HBsAg and HBeAg positivity rates across genotypes (A–F), as determined by the Abbott Architect assay. A 10-fold plasma dilution (10 µL in 100 µL of blocking buffer) reliably quantified (D) HBsAg and (E) HBeAg. Notably, HBeAg levels in samples tested from genotypes A and E were below the lower limit of detection (

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal α-HBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Infection, Blocking Assay

Journal: Pathogens (Basel, Switzerland)

Article Title: Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings.

doi: 10.3390/pathogens14030258

Figure Lengend Snippet: Figure 4. Comparison of various DNA isolation methods for determining the EC50 of TDF in HepAD38 cells. HepAD38 cells were seeded in 24-well plates or 96-well plates and treated with TDF for 14 days with media and drug replenishment every 72 h. (A) To determine the toxicity of TDF, HepAD38 cells in the 96-well plate format were incubated with Alamar Blue for 2 h prior to reading the absorbance at 560 nm and 600 nm. Data were normalized according to the manufacturer’s directions to determine cell viability. (B) HBeAg and HBsAg could be quantified from the supernatant of 96-well HepAD38 cells treated with Alamar Blue. Extracellular (cell supernatant, top panel) and intracellular (bottom panel) HBV DNA were determined in (C) 96-well and (D) 24-well plate formats and plotted as % HBV inhibition against log [TDF] to determine the EC50. To investigate if Alamar Blue negatively impacts EC50 determination, HepAD38 cells from the 96-well plate treated with Alamar Blue were also subjected to DNA isolation and quantification. (E) Summary of TDF EC50 values obtained from the treatment of HepAD38 cells in either a 24- or 96-well format. Assay Z′

Article Snippet: Primary antibodies used for antigen capture were mouse monoclonal α-HBsAg (Fitzgerald, 10-H05H) and mouse monoclonal α-HBeAg (Fitzgerald, 10-H10M).

Techniques: Comparison, DNA Extraction, Incubation, Inhibition

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The HBV large envelope protein initiates virion assembly by recruiting capsids at membrane rich domains related to late endosome

doi: 10.1007/s00018-025-05574-3

Figure Lengend Snippet: HBV core and HBV LHBs constructs. A: Core-NTD, linker (140–149) and CTD are depicted. eGFP or mCherry polypeptides have been inserted between positions 78 and 80. Y132A mutant is localized with a dark arrow. B: Diagram of a core dimer with an eGFP or mCherry positioned at the top of helix 3. C: Schematic representation of LHBs protein with preS1, preS2 and S domains. X indicates that AUG codons were changed to ACG to prevent translation of the M- and SHBs proteins. The eGFP and mCherry proteins were placed at the N-terminus of LHBs or SHBs. MD is the Matrix Domain that corresponds to the residues 103–124. D: Scheme of the interaction between the nucleocapsid labelled with eGFP and the MD domain of the i-PreS topology of LHBs labelled with mCherry. E: Western blot analysis of core, coreY132A, LHBs and L-ΔMD-HBs using anti-HBc (left panel) or anti-HBs (right panel). β–actin was used as control. F: Western blot analysis of mCherry, core-mCherry and mCherry-LHBs derivatives using anti-mCherry antibody. G: Western blot analysis of eGFP, core-eGFP and eGFP-LHBs derivatives using an anti-eGFP antibody. * corresponds to a partial hydrolysis of core-mCherry and core-mCherry derivatives as previously observed in

Article Snippet: The following primary antibodies were used: (i) a human anti-HBc polyclonal antibody at a 1:2000 dilution as described [ ], (ii) a mouse anti-HBs monoclonal antibody (1:2000, 70HG15, Biosynth), (iii) a rabbit polyclonal anti-GFP antibody (1:4000, ab6556, Abcam), (iv) a mouse Monoclonal anti-mCherry antibody [1C51] (1:4000, ab125096, Abcam), (v) and a rabbit polyclonal β-beta Actin antibody (1:4000, ab8227, Abcam).

Techniques: Construct, Mutagenesis, Western Blot, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The HBV large envelope protein initiates virion assembly by recruiting capsids at membrane rich domains related to late endosome

doi: 10.1007/s00018-025-05574-3

Figure Lengend Snippet: Co-localization of core-LHBs with markers of intracellular compartments. Huh-7 Cells were co-transfected with plasmids expressing core and LHBs. After 72 h, cells were fixed, permeabilized and immunostained. (A) In the green channel, images were obtained by immunofluorescence using anti-endosomal markers targeting endogenous proteins or using anti-HA for TSG101 or using aquamarine signal of LC3-Aquamarine protein in cyan. In yellow, the human anti-HBc polyclonal antibody targeting core, and in the magenta, the goat anti-HBs monoclonal antibody (Fitzgerald 70HG15) or rabbit anti-pre S1 R271 targeting LHBs protein. DAPI nuclear staining is shown in blue in the merge panel. The scale bars in white correspond to 10 µm. (B and C) The bar graphs show the mean values of the Pearson’s correlation coefficient (PCC) from 15–20 cells. The PCC was calculated independently for the green (or cyan) and yellow channels (B), or the green (or cyan) and magenta channels (C) for each cell. (D) After 72 h, cells transfected with core/core-mCherry (90/10) plasmids, or LHBs/ mCherry-LHBs (95/5) plasmids were washed and incubated with green lysoTracker in Leibovitz’s L-15 medium at 37 °C for 30 min. Images show the acidified vesicles staining in green, with core (upper panels) or LHBs (lower panels) in red. Cell periphery and nucleus are shown by orange dotted line. The scale bars in white correspond to 10 µm. (E) The mean values of the PCC of 20—25 cells are shown. In all cases scatter dots represent each cell analyzed from two independent experiments. The bars represent the mean values and the error bars the ± SD of the PCC. Representative images are shown

Article Snippet: The following primary antibodies were used: (i) a human anti-HBc polyclonal antibody at a 1:2000 dilution as described [ ], (ii) a mouse anti-HBs monoclonal antibody (1:2000, 70HG15, Biosynth), (iii) a rabbit polyclonal anti-GFP antibody (1:4000, ab6556, Abcam), (iv) a mouse Monoclonal anti-mCherry antibody [1C51] (1:4000, ab125096, Abcam), (v) and a rabbit polyclonal β-beta Actin antibody (1:4000, ab8227, Abcam).

Techniques: Transfection, Expressing, Immunofluorescence, Staining, Incubation