EpCAM expression of human malignant cells.

<p>(A) The mRNA of the malignant cell lines 5061, 5072 (pancreatic cancer), LNCAP, PC3 (prostate cancer), FemX-1, MEWO (melanoma), T47D, MCF7 (breast cancer), HT29 (colon cancer) and OH-1 (small cell lung cancer) were relatively quantified by qPCR, using GAPDH for normalization. 5072, LNCAP, PC3, T47D, MCF7, Caco2 and HT29 showed high expression levels of EpCAM mRNA. (B) EpCAM could be detected by Western blot analysis of HT29 cell lysate with a specific binding to antibody MOC31. Beta-actin was used as loading control. (C) EpCAM could positively be detected by flow cytometry analysis with MOC31 on all cancer cell lines, except of FemX-1 and MEWO. Isotype controls are shown as dotted lines.</p

Blood half life of mAbs T84.1 and IgG1.

<p>The blood half life of [¹²⁵I]-labelled T84.1 and nonspecific [¹²⁵I]-labelled IgG1 antibodies were determined in FemX-1 tumour bearing SCID mice. The blood half life of specific and control antibody were identical (T84.1 = 10.3 h (n = 4), IgG1 = 10.4 h (n = 4)).</p

CEACAM in vivo binding.

<p>[¹²⁵I]-Labelled T84.1 and nonspecific [¹²⁵I]-labelled IgG1 antibody were used for CEACAM <i>in vivo</i> binding in FemX-1 melanoma bearing SCID mice. There is a significant difference between specific and control antibody by FemX-1 melanoma, but not in other organs including blood (Two-way ANOVA, P<0.001, n = 6).</p

MRI of HT29 xenograft.

<p>HT29 colon carcinoma (a) grown at the subcutaneous injection site above muscles (b) appear hyper-intense in two-dimensional turbo spin-echo (TSE) sequence (MR images in axial orientation).</p

Evans Blue-Albumin complex distribution in vibratome sections of FemX-1 melanoma.

<p>Evans Blue-Albumin complex positive areas (blue) are recognizable at blood vessels in vital tumour tissue (a), at the tumour margin (c) and the transition (d) between vital tumour tissue and necrosis (b).</p

EpCAM <i>in vivo</i> binding.

<p>[125I]-Labelled specific anti EpCAM MOC31 and non-specific [125I]-labelled IgG1 antibody were used for EpCAM <i>in vivo</i> binding in HT29 carcinoma bearing SCID mice. There is a significant difference between specific and control antibody by HT29 carcinoma, but except for spleen and blood not in other organs (two-way ANOVA, P<0.001, n = 3). Standard deviations are indicated by bars.</p

IFP measurements of HT29 xenograft tumours.

<p>(A) Typical pressure recording of a HT29 tumour. After adjusting the pressure at 0 mbar and insertion of the needle (arrow a) in the tumour the pressure increased and decreased rapidly and reaches a stable value of 16 mbar. To test the fluid communication between pressure sensor and needle the tubing is compressed with a screw clamp (arrow b) which results in a sharp increase of the pressure, then an exponential decrease, followed by another period of stabilization before decompressing the tubing (arrow c) which produces a rapid decrease and an exponential increase. (B) Measurement of 6 different HT29 tumours showed a mean IFP of 11.9 (+/−4) mbar.</p

Summary of <i>in vitro</i> and <i>in</i> vivo CEACAM expression of all cell lines by different methods.

<p>Note that the highest CEACAM expression <i>in vitro</i> and <i>in vivo</i> showed the melanoma cell line FemX-1. The intensity (plus signs) and extent (percent) of the positive areas of 5 histological sections were determined by visual inspection of 3 independent observers.</p

CEACAM mRNA expression of human malignant cells.

<p>The mRNA of the malignant cell lines 5061, 5072 (pancreas), PC3, LNCAP (prostate), FemX-1, MEWO (melanoma), MCF7, T47D (breast), HT29 (colon) and OH-1 (small cell lung cancer) were relatively quantified by real time PCR, using GAPDH for normalization. Cell lines 5061, 5072, LNCAP and HT29 showed high expression levels of CEACAM mRNA.</p

CEACAM cell surface presence of tumour cells.

<p>CEACAM could positively be detected by FACS-analysis with mAb T84.1 by all cancer cell lines, except of T47D, MCF7 and PC3. Isotype controls are shown as dotted lines.</p