Cat. No.
MABL-2080
Application
in vivo, inhibition, ELISA
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
hu3B9
From
Recombinant Antibody
Specificity
This antibody is specific for human IL-4 and binds a non-contiguous epitope.
Alternative Names
IL4; BSF-1; Interleukin-4; B-cell stimulatory factor 1; Binetrakin; Lymphocyte stimulatory factor 1; Pitrakinra; 3B9
UniProt
P05112
Immunogen
The original antibody was generate by immunizing F1 hybrids of Balb/c and C57BL/6 with E. Coli-produced recombinant human IL-4.
Application Notes
The binding affinity of the original format and Fab fragment of this antibody were evaluated by ELISA. The original format of this antibody (mouse IgG1) exhibited a 0.2 nM Kd (~0.18 nM). The Kd of its Fab fragment was 0.3 nM. This antibody successfully inhibited IL-4 binding to its receptor, as evaluated using 125 I-rhIL4 binding to the gibbon cell line, MLA. This antibody successfully inhibited lymphocyte proliferation — the original format of this antibody inhibited 3 H-thymidine incorporation by human peripheral blood T lymphocytes stimulated with 133 pM IL-4 and human tonsillar B lymphocytes stimulated by 167 pM IL-4. The IC50 for inhibition of T cell proliferation was 30 pM, and for B cell proliferation 103 pM. The corresponding values for the Fab fragment were 108 and 393 pM. This antibody successfully inhibited CD23 induction — the original format of this antibody inhibited CD23 expression on human tonsil B lymphocytes stimulated with 8.3 pM IL-4 with an IC50 value of 136 pM. This antibody successfully inhibited IgE secretion — the original format and the Fab fragment of this antibody inhibited IgE secretion in the presence of 1.7 nM IL-4, giving IC50 values of 1.9 and 5.0 nM, respectively. The experiment was repeated using a lower concentration of IL-4, 667 pM, which reduced the IC50 value to 0.65 nM for the original format of the antibody. The humanized version of this antibody effectively inhibited the binding of IL-4 receptor-Fc fusion protein to IL-4 in a dose-dependent manner — at lower concentrations (10-20 nM), it achieved 50% inhibition, while at higher concentrations (1 µM), it achieves greater than 90% inhibition. In a pharmacokinetics experiment performed on male Sprague Dawley rats, the humanized version of this antibody exhibited low inter-animal variability with a slow plasma clearance value of 0.5 mL/h/kg and an approximate 11 days half-life with a biphasic disappearance pattern (US5914110A).
Antibody First Published
Note on publication
The original publication describes the generation and characterization of various anti-human IL-4 antibodies and their derivatives, including its humanization.
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.




