Identification of neo-vasculature markers and production of high-affinity recombinant antibodies for diagnostic and therapeutic applications

• Research unit: EU RFP
• Project number: 97.0091

Key words (English)

Two-dimensional gel electrophoresis; angiogenesis; phage antibodies; tumour targeting

Alternative project number (English)

EU project number: BIO4CT972149

Research programs (English)

EU-programme: 4. Frame Research Programme - 4.1 Biotechnology

Short description (English)

See abstract

Further information (English)

Full name of research-institution/enterprise:
ETH Zürich
Institut für Pharmazeutische Wissenschaften
ETH Hönggerberg, HCI G 396.4

Partners and International Organizations (English)

Coordinator: Università degli studi di Siena (I)

Abstract (English)

During the third year of the Project, our group has achieved the following results:
continuation of the use and validation of the ETH-2 antibody library
evaluation of the therapeutic potential of fusion proteins consisting of a recombinant human antibody specific for a marker of angiogenesis, fused to bioactive proteins (collaboration with the Genova group). In particular, the fusion protein between scFv(L19) and a truncated form of tissue factor has shown remarkable anti-tumour results in three different murine models of aggressive solid tumours. In our opinion, these experiments validate the design of the Project, in which we tried to combine the isolation of antibodies against markers of angiogenesis together with the investigation of their potential in diagnostic and therapeutic applications.
2D-PAGE study of proteins expressed (in different experimental conditions) by human endothelial cells and by human fibroblasts: two likely sources of markers of angiogenesis (collaboration with the Siena group and the Genova group). In particular, we have found proteins (such as collagen VI alpha I chain) which show strikingly different expression patterns in fibroblasts grown at pH 7.5 and pH 6.7.
Differential bio-panning of the ETH-2 library with proteins secreted by fibroblasts grown at pH 7.5 and pH 6.7. Isolation of three antibody clones, staining a novel protein, which is strongly expressed around new blood vessels in brain tumours but not in normal brain (collaboration with the Genova group).
Comparative analysis of different methods of 2D-gel staining (various Silver staining methods, Sypro Ruby staining with different fluorescent imaging modalities), together with an evaluation of the compatibility of these methods with mass spectrometric protein identification: an essential pre-requisite for the discovery of novel markers of angiogenesis (collaboration with the Siena group).

References in databases (English)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 97.0091