Anti-P. falciparum AMA1 Therapeutic Antibody ( 12Y-2) (CAT#: MRO-191CT)

Recombinant shark antibody specifically binds to Plasmodium falciparum AMA1, expressed in Chinese Hamster Ovary cells(CHO).

  • Published Data
  • Datasheet
  • MSDS
  • COA
ELISA

Figure 1 V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

ELISA comparison of binding of affinity purified proteins 12Y-1 (black) and 12Y-2 (grey) to lysozyme, AMA1, -amylase, and Tom70.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

ELISA

Figure 2 ELISA of V(NAR) 12Y-2 by error-prone PCR.

After three rounds of panning, 16 affinity-matured clones were tested for binding to AMA1 and to lysozyme (negative control antigen). The hatched line represents the parental (12Y-2) response level.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

SPR

Figure 3 The 12Y-2 triple-mutant shows parental affinity for AMA1.

BIAcore sensorgrams comparing the binding of identical concentrations of the parental (12Y-2), single mutant (14M-15), and triple mutant proteins to immobilized AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

SPR

Figure 4 Binding was measured at a constant flow rate of 5 ul/min with an injection volume of 35 ul. Dissociation was continued with HBS buffer until the response returned to the initial value before injecting the next sample

V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

SPR

Figure 5 Binding was measured at a constant flow rate of 5 ul/min with an injection volume of 35 ul. Dissociation was continued with HBS buffer until the response returned to the initial value before injecting the next sample

V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

SPR

Figure 6 IgNAR constant domain 1 is insufficient for dimer formation.

BIAcore sensorgrams comparing binding of proteins 17T-6 and 12Y-2 to immobilised AMA1 protein.

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

SPR

Figure 7 Dhlx-mediated dimerisation.

The peak-purified dimer fraction was analysed by BIAcore biosensor, compared to the parental 12Y-2

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

SPR

Figure 8 Tandem V(NAR)s in a single chain format

BIAcore sensorgrams comparing binding of proteins 12Y-2, 14M-15, and 15Q-1 to immobilised AMA1 protein.

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

ELISA

Figure 9 ELISA analysis of 40 12Y-2 variants binding to AMA-1 and a control negative antigen.

ELISA analysis of 40 12Y-2 variants binding to AMA-1 and a control negative antigen.

Inhib

Figure 10 Parasite Invasion Inhibition

Recombinant V(NAR) domains inhibit invasion of erythrocytes by P. falciparum 3D7 parasites. Data are represented as the mean ± standard error of quadruplicate experiments.

Henderson, K. A., Streltsov, V. A., Coley, A. M., Dolezal, O., Hudson, P. J., Batchelor, A. H., ... & Foley, M. (2007). Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition. Structure, 15(11), 1452-1466.

Inhib

Figure 11 Parasite Invasion Inhibition

Recombinant V(NAR) domains inhibit invasion of erythrocytes by P. falciparum 3D7 parasites. Data are represented as the mean ± standard error of quadruplicate experiments,except for P. falciparum W2mef parasites.

Henderson, K. A., Streltsov, V. A., Coley, A. M., Dolezal, O., Hudson, P. J., Batchelor, A. H., ... & Foley, M. (2007). Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition. Structure, 15(11), 1452-1466.


Specifications

  • Host Species
  • Shark
  • Species Reactivity
  • P. falciparum
  • Clone
  • 12Y-2
  • Applications
  • ELISA, SPR, Inhib

Applications

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

Target

  • Alternative Names
  • apical membrane antigen 1; AMA1; AMA-1; Plasmodium falciparum; P. falciparum; Pf83; RMA-1; RMA1

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 "AMA1"

* 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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