Cat. No.
MABL-3931
Application
Biacore, WB, ELISA, FC, IF, IHC
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
L8A4
From
Recombinant Antibody
Specificity
Binds EGFRvIII, a mutated form of EGFR, plays a prominent role in tumorigenesis.
Alternative Names
Epidermal Growth Factor Receptor variant III
UniProt
P00533
Immunogen
The original antibody was isolated from a yeast-display scFv library was constructed by homologous recombination of the thrombotic thrombocytopenic purpura (TTP) patient repertoire originally expressed on M13 bacteriophage in the novel vector pAGA2 for yeast-display expression. The library was panned against human TEM1 recombinant protein.
Application Notes
The in vitro binding characterization of this antibody was done using ELISA and flow cytometry. This antibody can recognize TEM1-expressed on mammalian cells, like human or murine cell lines expressing TEM1 endogenously (HEK-293T, MOV1 and 2H11) or stably transduced with pHRSINTEM1 (hTEM1-MS1, hTEM1-H5V and hTEM1-SKOV3) in a flow cytometric assay. This antibody binds specifically to Endosialin/TEM1-expressing ovarian tumor in vivo, with functional stability over 48 hrs. (PMID: 20837020). The original scFv version of this antibody binds human TEM-1 with a dissociation constant of Kd= 2 nM (PMID: 29313073).The scFv78-saporin immunoconjugate generated using this antibody exerted dose- dependent cytotoxicity with high specificity to TEM1-positive cells in vitro (PMID: 27933426). This antibody was used in the generation of a multivalent Fc-fusion protein called 78Fc, which has desirable characteristics for immuno-imaging applications.
Antibody First Published
Zhao et al. Rapid isolation of high affinity human antibodies against the tumor vascular marker Endosialin/TEM1, using a paired yeast display/secretory scFv library platform. J Immunol Methods. 2011 Jan 5; 363(2): 221–232. PMID:20837020
Note on publication
Describes the generation of a recombinant antibody (scFv) library derived from the repertoire of a patient with thrombotic thrombocytopenic purpura (TTP) and isolation of an anti-TEM1 antibody.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
