Expression of a bispecific dsFv-dsFv' antibody fragment in Escherichia coli.
A bispecific disulfide-stabilized Fv antibody fragment (dsFv-dsFv') consisting of two different disulfide-stabilized Fv antibody fragments connected by flexible linker peptides was produced by secretion of three polypeptide chains into the periplasm of Escherichia coli. The dsFv-dsFv' molecules were enriched by immobilized metal affinity chromatography and further purified by anion-exchange chromatography. The recombinant antibody constructs retained the two parental antigen binding specificities and were able to cross-link the two different antigens. The described dsFv-dsFv' design might be of particular value for therapeutic in vivo applications since improved stability is expected to be combined with minimal immunogenicity.
Schmiedl
A
A
Universität Heidelberg, Institut für Molekulare Genetik, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany.
Breitling
F
F
Dübel
S
S
D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000915:Antibody Affinity; D004220:Disulfides / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology*; D005822:Genetic Vectors / Q000138:chemical synthesis; D015151:Immunoblotting; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D051379:Mice; D016254:Mutagenesis, Insertional; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
D018033:Antibodies, Bispecific; D004220:Disulfides; D007135:Immunoglobulin Variable Region; D010455:Peptides; D011994:Recombinant Proteins