Recombinant Mouse Anti-Horse Equ c 1 Antibody (mAb220)

Natural Equ c 1 (10 μg/ml) was coated on the microtitration plate, and biotinylated mAbs (1 μg/ml) (*) as indicated were co-incubated with competitor mAb 118 (open bar), mAb 197 (hatched bar), or mAb 220 (closed bar) at 10 μg/ml. Results are expressed as the percentage of residual biotinylated mAb binding revealed by peroxidase-labeled streptavidin.

Lascombe, M. B., Grégoire, C., Poncet, P., Tavares, G. A., Rosinski-Chupin, I., Rabillon, J., ... & Alzari, P. M. (2000). Crystal Structure of the Allergen Equ c 1 A DIMERIC LIPOCALIN WITH RESTRICTED IgE-REACTIVE EPITOPES. Journal of Biological Chemistry, 275(28), 21572-21577.

Equ c 1 (10 μg/ml), either natural (A) or recombinant (B), was coated on a microtitration plate, and IgE serum representative of eight allergic patients (1/10 dilution) was co-incubated with varying concentrations of competitor antibody: mAb 118 (○), mAb 197 (▵), mAb 220 (■), mAb 65 (⋄), or rabbit anti-Equ c 1 polyclonal Abs (●). Binding of human IgE was detected with biotinylated goat anti-human IgE Abs followed by peroxidase-coupled streptavidin and revealed with o-phenylene diamine according to the manufacturer's recommendations. Results are expressed as the percentage of inhibition of human IgE binding.

Lascombe, M. B., Grégoire, C., Poncet, P., Tavares, G. A., Rosinski-Chupin, I., Rabillon, J., ... & Alzari, P. M. (2000). Crystal Structure of the Allergen Equ c 1 A DIMERIC LIPOCALIN WITH RESTRICTED IgE-REACTIVE EPITOPES. Journal of Biological Chemistry, 275(28), 21572-21577.

The electrophoresis and blotting was performed using 10 ml of bacterial supernatant (diluted 1:500). Immunological detection was performed using monoclonal antibodies mAb 118 (lane 1), mAb 197 (lane 2), mAb 220 (lane 3), a polyclonal mouse serum (lane 4), and polyclonal rabbit serum (lane 6) raised against HD Equ c1 and using IgE from human serum (representative of 16 tested human sera) of a patient suffering from allergic reactions to horses (lane 8). Nonimmunized mouse (lane 5) and rabbit (lane 7) sera and human serum from nonallergic patients (lane 9) were used as controls.

Gregoire, C., Rosinski-Chupin, I., Rabillon, J., Alzari, P. M., David, B., & Dandeu, J. P. (1996). cDNA cloning and sequencing reveal the major horse allergen Equ c1 to be a glycoprotein member of the lipocalin superfamily. Journal of Biological Chemistry, 271(51), 32951-32959.