CAMELID-VNH: Structure, properties and biotechnological applications o VHH antibody fragments from camelids

• Forschungsstelle: EU FRP
• Projektnummer: 98.0214
• Projekttitel Englisch: CAMELID-VNH: Structure, properties and biotechnological applications o VHH antibody fragments from camelids

Schlüsselwörter (Englisch)

Recombinant antibodies; protein engineering; protein design; molecular biotechnology

Alternative Projektnummern (Englisch)

EU project number: BIO4CT980048

Forschungsprogramme (Englisch)

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

Kurzbeschreibung (Englisch)

See abstract

Partner und Internationale Organisationen (Englisch)

Coordinator: VIB-VUB, Bruxelles

Abstract (Englisch)

The naturally occuring antibodies devoid a light chain and of a CH1 domain discovered earlier in camelids constitute a valuable tool for various applications. To successfully implement this new technology, fundamental knowledge on the structural and molecular characteristics of the camelid VHH antibody fragments is required. Especially, knowledge on the structural and functional stability during production, storage and application conditions is essential. For example, many consumer products are subjected to a heat treatment during the packaging process, which requires heat stability of the antibody fragments. Alternatively , during storage and application the fragments might be exposed to high concentrations of detergents and/or extreme pH or ionic strength. e.g. in toothpaste or detergents. Antibody fragments need to stable over the shelve life period of the product and under application conditions.
The aim of this project was to obtain in depth knowledge on the structural and functional stability relationships of the camelid VHH antiobody fragments to be able to design VHH mutants with improved stability with respect to high temperatures and surfactants and to obtain higher expression levels in heterologuous organisms.
Two different research routes have been explored to these goals. The first one was to obtain insight on the temperature stability of the llama VHH fragments. To this end the temperature unfolding of a fragment has been investigated in detail. This knowledge will be applied in mutagenesis studies to improve the temperature stability ofthe llama VHH antibody fragments. The second route was to obtain insight on the stability of these fragments at application conditions. The results obtained in the initial studies have led to the design of a first generation of variants. Unfortunately, these variants did not show the expected enhanced stability towards surfactants. However, new ideas have been generated which will be further tested to obtain VHH fragments with improved stability and expression levels

Datenbankreferenzen (Englisch)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 98.0214