Global fitted (dotted line) SPR data of murine MX35 and Rebmab200 binding to immobilized synthetic NaPi2b epitope.

<p>MX35 (A) and Rebmab200 (B) were injected at concentrations ranging from 5 to 80 nM. After a 10 min association phase, the dissociation phase was followed for additional 10 min. Following double subtractive referencing, the curves were plotted using a 1∶1 Langmuir binding model, using Biacore T100 Evaluation Software. The solid line represents the experimental data and the dotted line the mathematical model for the binding of MX35 and Rebmab200 to the synthetic NaPi2b epitope.</p

Comparison of ADCC activity between MX35 and Rebmab200 (stable pool) over a range of mAb concentrations.

<p>The % of cytotoxicity represents antibody-mediated cell lysis measured by release of ⁵¹Cr from labeled ovarian cancer cells (OVCAR-3). Effector cells were obtained from donated human peripheral blood. Rebmab100 was used as a positive control, and Zenapax (Roche) was used as a negative control. The assay was repeated with MCF-7 cells, a NaPi2b negative and Le^Y positive (Rebmab100 antigen) tumor cell line, showing no ADCC results (data not shown).</p

Immunoreactivity of MX35 and Rebmab200 in live cells by flow cytometry.

<p>Immunoreactivity of MX35 and Rebmab200 in live cells by flow cytometry.</p

Immunoglobulin HC and LC mRNA levels and productivity of the most representative Rebmab200 clones as determined by RT-qPCR.

<p><b>GAPDH was used as a reference gene.</b> Vector represents PER.C6®cells transfected with mock vector; PER.C6® corresponds to the parental cell. Error bars represent the standard deviation of triplicates. Qp represents specific productivity levels.</p

Immunoreactivity of MX35 and Rebmab200 in tumor tissues by immunohistochemistry.

<p>Immunoreactivity of MX35 and Rebmab200 in tumor tissues by immunohistochemistry.</p

Comparison of the immuno-affinity of MX35 (left) at 1/50 dilution and Rebmab200 (right) at 1/100 dilution in parallel reactions using the same buffers and amplification and development conditions.

<p>Serial sections of serous papillary ovary adenocarcinoma (A), clear cell renal carcinoma (B), large cell poorly differentiated carcinoma of the lung (C) and invasive ductal carcinoma of the breast (D). Original magnification = 100×.</p