Recombinant Human Anti-RSV Antibody (AM14)
CAT#: PABL-321
Recombinant Human Antibody (AM14) 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. This antibody potently neutralized laboratory strains and clinical isolates of RSV from both A and B subtypes.
Tested Data
Published Data
SDS-PAGE
Figure 1 Recombinant Human Anti-RSV Antibody (PABL-321) in SDS-PAGE
SDS-PAGE analysis of PABL-321 in non-reduced (Lane 2) and reduced (Lane 1) conditions.
SEC-HPLC
Figure 2 Recombinant Human Anti-RSV Antibody (PABL-321) in HPLC
The purity of PABL-321 was greater than 95% as determined by SEC- HPLC.
WB
Figure 3 Recombinant Human Anti-RSV Antibody (PABL-321) in WB
Western blot analysis of PABL-321 was performed by loading Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-321 and HRP Goat Anti-Human IgG as a secondary antibody. Chemiluminescent detection was performed.
Lane 1: Reduced antigen (1 μg)
DB
Figure 4 Recombinant Human Anti-RSV Antibody (PABL-321) in DB
Dot Blot analysis of PABL-321 was performed by coating with Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag).
PABL-321 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG as a secondary antibody (1: 2000)
ELISA
Figure 5 Recombinant Human Anti-RSV Antibody (AM14) (PABL-321) in ELISA
ELISA analysis of PABL-321 was performed by coating with Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag).
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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 Neutralization of laboratory strains and clinical isolates of RSV.
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 AM14 to immobilized RSV F proteins was measured using a Luminex system.
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 Binding of AM14 Fab to immobilized prefusion RSV F was measured by surface plasmon resonance. Best fit of the data to a 1:1 binding model is shown in red.
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 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 6 AM14 is specific for cleaved, trimeric RSV F.
Binding of antibodies AM14 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 7 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.
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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
- AM14
- 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 ELISA and Neutralization. For details, refer to Published Data.
Target
- Alternative Names
- Human respiratory syncytial virus; RSV
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Customer Review
There are currently no Customer reviews or questions for PABL-321. Click the button above to contact us or submit your feedback about this product.
Laura Bennett05.Jun,23
Excellent for WB
I was quite happy with AM14's ability to neutralize RSV in my cell culture tests. The only reason I'm giving it four stars is due to the shipping delay, but the antibody is excellent.
Alex Thompson18.May,21
precise RSV detection
This antibody is one of the most effective we have used to detect RSV. It has excellent specificity and sensitivity. The signal is robust and clear, greatly improving the sensitivity of the test. This reliability allows us to focus more on our research without worrying about unpredictability and non-specific binding. I would definitely buy it again.
Jason Lee08.Oct,23
Highly Dependable for ELISA
Overall, a great antibody for detecting RSV. It provided reproducible results in ELISA. The datasheet could use a bit more detail, but that's a minor issue. Highly recommended otherwise.
Dor***y11.Sep,23
Effective and Reliable for RSV Detection
We have been utilizing the Recombinant Human Anti-RSV Antibody (AM14) (CAT#: PABL-321) from Creative Biolabs in our RSV detection assays, and the results have been exceptional. The antibody demonstrates excellent specificity and sensitivity for RSV, allowing precise detection of the virus in clinical samples. The ELISA assays have consistently yielded reproducible results with minimal background interference.
Q&As
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What is the main application of the Recombinant Human Anti-RSV Antibody (AM14)?
A: The Recombinant Human Anti-RSV Antibody (AM14) is primarily used for the detection and neutralization of Respiratory Syncytial Virus (RSV) in various types of assays, including ELISA, Western blotting.
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How is the specificity of the Recombinant Human Anti-RSV Antibody (AM14) validated?
A: The antibody's specificity is verified by testing it against RSV-infected samples and control groups to ensure that it binds only to RSV antigens and does not cross-react with other viral proteins.
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Does AM14 cross-react with other respiratory viruses, and if so, could this lead to off-target effects?
A: AM14 has been thoroughly studied for off-target binding and demonstrates good specificity for RSV with limited cross-reactivity to other common respiratory viruses, hence reducing possible off-target consequences.
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Can the Recombinant Human Anti-RSV Antibody (AM14) be used for quantitative assays?
A: Yes, the antibody can be used for quantitative assays such as ELISA. Proper calibration with known standards is required to achieve accurate quantification.
View the frequently asked questions answered by Creative Biolabs Support.
Cite This Product
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# PABL-321, RRID: AB_3111648)
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Downloadable Resources
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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.
Datasheet
MSDS
COA
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