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

• Unité de recherche: PCRD EU
• Numéro de projet: 97.0091
• Titre du projet anglais: Identification of neo-vasculature markers and production of high-affinity recombinant antibodies for diagnostic and therapeutic applications

Mots-clé (Anglais)

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

Autre Numéro de projet (Anglais)

EU project number: BIO4CT972149

Programme de recherche (Anglais)

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

Description succincte (Anglais)

See abstract

Autres indications (Anglais)

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

Partenaires et organisations internationales (Anglais)

Coordinator: Università degli studi di Siena (I)

Résumé des résultats (Abstract) (Anglais)

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).

Références bases de données (Anglais)

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