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Single-chain Fv fragments exhibiting monovalent binding can be converted into bivalent molecules by reducing the linker connecting the two variable domains to approximately five residues resulting in homodimerization of two polypeptide chains. This protocol describes the cloning steps to convert an scFv into a diabody molecule and provides some information for production and analysis.
KeywordsAntibody Fragment SacI Site Calf Intestine Alkaline Phosphatase Antibody Sequence Restriction Site Analysis
- Kortt A, Lah M, Oddie GW, Gruen CL, Burns JE, Pearce LA, Atwell JL, McCoy AK, Howlett GJ, Metzger DW, Webster RG, Hudson PJ (1997) Single-chain Fv fragments of anti-neuraminidase antibody NC10 containing five- and ten-residue linkers form dimers and with zero-residue linker a trimer. Protein Eng 10:423–433PubMedCrossRefGoogle Scholar