Recombinant Human Anti-RSV Antibody (D25) (CAT#: PABL-322)
Recombinant Human Antibody (D25) is capable of binding to RSV, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-RSV mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-RSV mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.
Specific Inquiry
Figure 1 Internalization of Fab fragments.
RSV-infected HEp-2 cells were incubated with RSV F-specific MAbs D25, AM14, 5C4, and MPE8 or corresponding monomeric Fab fragments at the same concentration for 90 min to induce internalization. Afterwards the cells were fixed, permeabilized, and stained with AF488 human anti-goat IgG or AF488 chicken anti-mouse IgG (green). Nuclei were visualized with DAPI (blue). The amount of internalized vesicles was quantified in 50 positive cells.
Leemans, A., De Schryver, M., Van der Gucht, W., Heykers, A., Pintelon, I., Hotard, A. L.,... & Broadbent, L. (2017). Antibody-induced internalization of the human respiratory syncytial virus fusion protein. Journal of virology, 91(14), e00184-17.
Figure 2 N426D disrupts binding of AM14 to prefusion F. Relative binding of D25, 101F, MPE8 and AM14 to cell surface-expressed prefusion F (grey) or prefusion F containing the AM14 escape mutation, N426D (white) was measured by flow cytometry. Data were normalized to motavizumab binding. Binding of D25, 101F and MPE8 to N426D was comparable to wild-type prefusion F, whereas AM14 binding to N426D was reduced four-fold.
Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.
Figure 3 Binding of antibodies (A) AM14, (B) D25 to uncleaved monomeric RSV F (open black triangles), cleaved monomeric RSV F (black circles), uncleaved postfusion RSV F (open blue triangles), cleaved postfusion RSV F (blue circles), uncleaved prefusion RSV F (open red triangles) and cleaved prefusion RSV F (red circles) was measured by ELISA.
Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.
Figure 4 AM14 stabilizes RSV F trimer in the absence of the foldon trimerization motif. Size-exclusion chromatography profiles from a Superose 6 column are shown for AM14 Fab or D25 Fab complexed with prefusion RSV F containing the foldon trimerization motif (black and grey, respectively) and for AM14 Fab or D25 Fab co-expressed with RSV F ectodomain without foldon (red and blue, respectively).
Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.
Figure 5 RSV neutralization by antibodies. Palivizumab is the FDA-approved prophylactic antibody that prevents severe RSV disease.
McLellan, J. S., Chen, M., Leung, S., Graepel, K. W., Du, X., Yang, Y.,... & Kumar, A. (2013). Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science, 340(6136), 1113-1117.
Figure 6 ELISA measuring antibody binding to postfusion F glycoprotein.
McLellan, J. S., Chen, M., Leung, S., Graepel, K. W., Du, X., Yang, Y.,... & Kumar, A. (2013). Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science, 340(6136), 1113-1117.
Figure 7 Highly effective RSV-neutralizing antibodies target a site at the membrane-distal apex of the prefusion F trimer.
(A) The ability of antibodies to block D25 binding to RSV-infected cells was measured as a function of antibody concentration. (B) Analysis of RSV F-Fab complexes by negative stain electron microscopy: Reprojection of a 12 Å slice through the crystal structure of RSV F + D25 Fab filtered to 10 Å resolution (left). A slice was used to emphasize visibility of the F glycoprotein cavity. Aligned average of 263 particles of RSV F + D25 Fab (middle left). Aligned average of 550 particles of RSV F + AM22 Fab (middle -right). Aligned average of 171 particles of RSV F + 5C4 Fab (right). Scale bar is 50 Å. (C) Fusion inhibition and (D) attachment inhibition activity for antibodies targeting antigenic site Ø and F-specific antibodies targeting other antigenic sites. For the attachment-inhibition assay, heparin was used as a positive control.
McLellan, J. S., Chen, M., Leung, S., Graepel, K. W., Du, X., Yang, Y.,... & Kumar, A. (2013). Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science, 340(6136), 1113-1117.
Specifications
- Immunogen
- The details of the immunogen for this antibody are not available.
- Host Species
- Human
- Derivation
- Human
- Type
- IgG
- Specificity
- Tested positive against native RSV
- Species Reactivity
- RSV
- Clone
- D25
- Applications
- WB, ELISA, Neut, FuncS
Product Property
- Purity
- >95% by SDS-PAGE and HPLC analysis
- Storage
- Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.
Applications
- Application Notes
- The antibody was validated for Neutralization, ELISA and Function Assay. For details, refer to Published Data.
Target
- Alternative Names
- RSV; Human respiratory syncytial virus
Related Resources
Tang, Wei, et al. "Mechanism of cross-resistance to fusion inhibitors conferred by the K394R mutation in respiratory syncytial virus fusion protein." Journal of Virology 95.20 (2021): 10-1128.
This study investigates the mechanism of cross-resistance to fusion inhibitors conferred by the K394R mutation in the respiratory syncytial virus (RSV) fusion protein. The researchers employed a dual-luciferase protocol to discover LF-6, a small-molecule inhibitor that targets the RSV fusion glycoprotein. They found that the K394R mutation not only conferred resistance to LF-6 but also to other RSV fusion inhibitors, including those in clinical development. This mutation destabilizes the F protein and enhances its membrane fusion activity, increasing viral resistance. The study highlights the need for alternative strategies to combat RSV infections, given the resistance risks posed by the K394R mutation.
In this research, Creative Biolabs provided crucial reagents that facilitated the experiments. Specifically, the D25 monoclonal antibody (Cat# PABL-322) and Motavizumab (Cat# TAB-709) were used for detecting RSV F protein in various assays, including the immunofluorescence and cell-surface triggering assays. These antibodies were essential for accurately identifying the effects of mutations on the RSV F protein and assessing the resistance mechanisms. The products from Creative Biolabs significantly contributed to the detailed analysis and validation of the findings related to the K394R mutation and its impact on RSV fusion inhibitor resistance.
Gilbert, Brian E., et al. "Respiratory syncytial virus fusion nanoparticle vaccine immune responses target multiple neutralizing epitopes that contribute to protection against wild-type and palivizumab-resistant mutant virus challenge." Vaccine 36.52 (2018): 8069-8078.
This study focused on the development and evaluation of an RSV F nanoparticle vaccine targeting human respiratory syncytial virus (RSV), which is a major cause of severe respiratory infections in various vulnerable populations, including newborns and the elderly. The RSV F nanoparticle vaccine was designed to induce a broad polyclonal antibody response against multiple neutralizing epitopes on the fusion (F) protein of RSV. These antibodies were shown to neutralize both wild-type RSV and palivizumab-resistant mutant strains in animal models, particularly in cotton rats. The vaccine's efficacy was enhanced with the use of an aluminum phosphate adjuvant, which significantly increased the antibody avidity and protection against RSV challenges.
Creative Biolabs contributed to this study by providing the monoclonal antibody D25 (Cat#: PABL-322), which was used for competitive binding assays to characterize the antibody responses elicited by the RSV F nanoparticle vaccine. The D25 antibody specifically targets antigenic site Ø on the pre-fusion F conformation of the RSV F protein. The inclusion of this antibody was crucial for demonstrating that the vaccine-induced antibodies could bind to conserved epitopes on both pre-fusion and post-fusion F conformations, supporting the vaccine's potential to provide broad protection against RSV.
More Infomation
Product Notes
This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:
• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
See more details about Hi-Affi™ recombinant antibody benefits.
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| CAT | Product Name | Application | Type |
|---|---|---|---|
| TAB-098 | Anti-RSV Recombinant Antibody (Felvizumab) | IF, IP, Neut, FuncS, ELISA, FC, ICC | IgG1 - kappa |
Customer Reviews and Q&As
Customer Review
Q&As
-
Excellent for RSV Research
The Anti-RSV Antibody has been excellent for our RSV detection assays. Its specificity and sensitivity are top-notch, providing clear and reliable results. This antibody has significantly improved the accuracy of our experiments and saved us a lot of troubleshooting time. -
Reliable in Immunoassays
We incorporated this antibody into our immunoassays, and it has been a great asset. The specificity for RSV is precise, allowing for accurate detection and analysis. The reliable performance has made it an invaluable tool in our lab. -
Consistent High-Quality Results
This antibody has proven to be consistent and reliable in our assays. The high-quality, reproducible results have significantly improved our experiments, making it a crucial part of our research toolkit. The strong affinity and specificity for RSV have been particularly beneficial.
Cite This Product
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# PABL-322, RRID: AB_3111649)
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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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