New protein engineering approaches to multivalent and bispecific antibody fragments.

Multivalency is one of the hallmarks of antibodies, by which enormous gains in functional affinity, and thereby improved performance in vivo and in a variety of in vitro assays are achieved. Improved in vivo targeting and more selective localization are another consequence of multivalency. We summarize recent progress in engineering multivalency from recombinant antibody fragments by using miniantibodies (scFv fragments linked with hinges and oligomerization domains), spontaneous scFv dimers with short linkers (diabodies), or chemically crosslinked antibody fragments. Directly related to this are efforts of bringing different binding sites together to create bispecific antibodies. For this purpose, chemically linked fragments, diabodies, scFv-scFv tandems and bispecific miniantibodies have been investigated. Progress in E. coli expression technology makes the amounts necessary for clinical studies now available for suitably engineered fragments. We foresee therapeutic advances from a modular, systematic approach to optimizing pharmacokinetics, stability and functional affinity, which should prove possible with the new recombinant molecular designs.