title: A comparison of strategies to stabilize immunoglobulin Fv-fragments.
abstract: Fv-Fragments of antibodies may dissociate at low protein concentrations and are too unstable for many applications at physiological temperatures. To stabilize Fv-fragments against dissociation, we have tested and compared three different strategies on the Fv-fragment of the well-characterized phosphocholine binding antibody McPC603 expressed and secreted in Escherichia coli: chemical cross-linking of the variable domains, introduction of an intermolecular disulfide bond, and construction of a peptide linker to produce a "single-chain" Fv-fragment. All the linked fragments show hapten affinities nearly identical with that of the whole antibody independent of protein concentration and are significantly (up to 60-fold) stabilized against irreversible thermal denaturation. All genetically engineered linked Fv-fragments can be obtained in native conformation in E. coli. The reported strategies for generating Fv-fragments with improved physicochemical properties may extend their usefulness in biotechnology as well as in therapeutic and diagnostic applications.
authors.affiliation: Genzentrum, Universität Müchen, Max-Planck-Institut für Biochemie, Martinsried, FRG.
authors.forename: R
authors.initials: R
authors.lastname: Glockshuber
authors.forename: M
authors.initials: M
authors.lastname: Malia
authors.forename: I
authors.initials: I
authors.lastname: Pfitzinger
authors.forename: A
authors.initials: A
authors.lastname: Plückthun
mesh_terms: D055598:Chemical Phenomena; D002621:Chemistry; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004220:Disulfides; D004926:Escherichia coli / Q000378:metabolism; D005453:Fluorescence; D005976:Glutaral / Q000494:pharmacology; D006358:Hot Temperature; D007128:Immunoglobulin Fragments; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D004220:Disulfides; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D005976:Glutaral