Anti-Influenza A Virus H1N1 Hemagglutinin HA1 subunit Therapeutic Antibody (R2b-E8) (CAT#: MRO-231LC-VHH)

This product is a recombinant alpaca anti-influenza A virus H1N1 single domain antibody. R2b-E8 specifically binds to hemagglutinin HA1 subunit and can be potentially usd in the treatment of influenza virus infection.

  • Published Data
  • Datasheet
  • MSDS
  • COA
FC

Figure 1 Display of hemagglutinin on yeast and evaluation of binding of single domain antibodies.

FACS plots of seven HA specific single domain antibodies (sdAbs) R1a-F5, R1a-G6, R2b-E8, R2b-D9, R1a-A5, R1a-B6, R2a-G8 binding to yeast displayed HA0. Negative controls sdAb R1a-G2 and no sdAb control are shown. The vertical arrow indicates absence of binding.

Gaiotto, T., & Hufton, S. E. (2016). Cross-neutralising nanobodies bind to a conserved pocket in the hemagglutinin stem region identified using yeast display and deep mutational scanning. PloS one, 11(10), e0164296.

ELISA

Figure 2 Specificity of single domain antibodies to different influenza antigen reference reagents.

ELISA comparing binding of purified VHH antibodies at 30 mg/ml against H1N1, seasonal H1N1 and H5N1.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

SPR

Figure 3 Antibody affinity on recombinant HA by surface plasmon resonance.

Affinity on recombinant H1-HA, H1N1.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

FuncS

Figure 4 Viral neutralisation activity of cross-reactive sdAbs.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

ELISA

Figure 5 Characterisation of antibody epitopes.

ELISA showing reactivity of purified antibodies at 30 mg/ml to HA antigen standard H1N1 either treated with low pH or neutral pH.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

ELISA

Figure 6 Characterisation of antibody epitopes.

ELISA showing binding of phage displayed HA gene fragments to purified sdAbs.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

SPR

Figure 7 Grouping of antibody epitopes.

SPR co-injection experiments were used to determine if pairs of VHH antibodies can bind to recombinant H1-HA simultaneously. The antibodies R1a-A5, R1a-B6, R2b-D9, R2b-E8, R1a-C5 and R2a-G9 were tested in combination with each other. Example series of sensorgrams of R2b-D9 injected as the first antibody followed by injection of each of the other antibodies as indicated R2b-D9/R2b-E8.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

SPR

Figure 8 Grouping of antibody epitopes.

SPR co-injection experiments were used to determine if pairs of VHH antibodies can bind to recombinant H1-HA simultaneously. The four antibodies R2b-D9, R1a-B6, R2b-E8 and R1a-A5 predicted to share overlapping epitopes were further analysed by injecting each VHH on their own or as equimolar mixture of all four antibodies (Mixture).

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

SPR

Figure 9 Grouping of antibody epitopes.

The sensorgrams indicate the Rmax value for each of the individual antibodies with no significant increase in Rmax following injection of the antibody mixture, which suggests these antibodies recognise an overlapping epitope or an epitope that hinders binding of a second antibody. If the antibodies recognised non-overlapping or non-hindering sites the Rmax would be expected to be the sum of the individual Rmax values. This was confirmed when R1a-C5 which was predicted to recognise a non-overlapping epitope was included in a equimolar mixture of five antibodies and the response was seen to increase by an amount equivalent to R1a-C5 binding individually.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

SPR

Figure 10 Evaluation of competition with C179.

The antibody C179 was injected first and the test VHH antibody injected as the second antibody species C179/R2b-E8. The solid line represents co-injection of a first antibody and then a second antibody whereas a dotted line represents injection of a first antibody followed by injection of buffer.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.


Specifications

  • Host Species
  • Alpaca
  • Derivation
  • Phage display library
  • Type
  • Single domain antibody
  • Specificity
  • Influenza A virus H1N1
  • Clone
  • R2b-E8
  • Applications
  • ELISA, FuncS, SPR, FC
  • Related Disease
  • Influenza virus infections

Applications

  • Application Notes
  • The antibody was validated for ELISA, Functional Assay, Surface plasmon resonance, Flow Cytometry. For details, refer to Published Data.

Target

  • Alternative Names
  • Influenza A Virus H1N1 Hemagglutinin HA1 subunit

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.

See other products for "Influenza A Virus H1N1 Hemagglutinin HA1 subunit"

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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