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Transgenic antibodies : development of an integrated vector system for the expression of immunoglobulinsin different cellular compartments of mammaliancells


Many products of major therapeutic value can be obtained only from animal cells grown in culture. The aim of this project is to improve the processes currently employed in this sector of biotechnology with special emphasis on the vectors and promoters used and on the quality of post translational modification.

It is anticipated that in the future, monoclonal antibodies will be routinely isolated from phage libraries. Similarly, the expression of binding domains derived from antibodies within different cellular compartments of mammalian cells is likely to become a widely used technique to inhibit the function of recognised molecules. An important feature of the phage antibody technology is that the process of antibody selection is simultaneous with that of the cloning of the corresponding encoding sequences.

It would, therefore, be highly desirable that vectors for the expression of antibody domains within mammalian cells should be compatible with the phage vectors used to select antibody specificities. The development of such an integrated vector system for the expression of immunoglobulin domains within different cellular compartments of mammalian cells after they have been selected from phage antibody libraries is the objective of this project.

By the end of this project we would expect a researcher to be able to screen a phage library of diversity large enough to allow the selection of a monoclonal phage antibody (as ScFv of FAb) which recognises the antigen used for screening at a useful affinity. The cloning of the antibody binding specificity into an appropriate vector using convenient and consistent sites would allow expression of the selected antibody in the most suitable form (as ScFv, FAb, or complete antibody) within the particular cellular compartment of interest of a mammalian cell (intracytoplasmic, nuclear, secretory, endoplasmic reticulum), by the incorporation of appropriate protein tags at N or C terminal ends. The use of different promoters (tissue specific, inducible, constitutive) and vectors (plasmid, retroviral) will allow further refinement of expression. By this means the expression of targeted molecules is expected to be inhibited in situ.

Funding Scheme

CSC - Cost-sharing contracts


Società Italiana per la Ricerca Scientifica Srl
Via Giovanni Paisiallo 47/ C
00198 Roma

Participants (2)

Cambridge Antibody Technology Ltd.
United Kingdom
The Science Park Melbourn
SG8 6EJ Royston
Via Beirut 2-4
34014 Grignano