Properties of a single-chain antibody containing different linker peptides.
Single-chain antibodies were constructed using six different linker peptides to join the VH and VL domains of an anti-2-phenyloxazolone (Ox) antibody. Four of the linker peptides originated from the interdomain linker region of the fungal cellulase CBHI and consisted of 28, 11, six and two amino acid residues. The two other linker peptides used were the (GGGGS)3 linker with 15 amino acid residues and a modified IgG2b hinge peptide with 22 residues. Proteolytic stability and Ox binding properties of the six different scFv derivatives produced in Escherichia coli were investigated and compared with those of the corresponding Fv fragment containing no joining peptide between the V domains. The hapten binding properties of different antibody fragments were studied by ELISA and BIAcoreTM. The interdomain linker peptide improved the hapten binding properties of the antibody fragment when compared with Fv fragment, but slightly increased its susceptibility to proteases. Single-chain antibodies with short CBHI linkers of 11, six and two residues had a tendency to form multimers which led to a higher apparent affinity. The fragments with linkers longer than 11 residues remained monomeric.
Alfthan
K
K
VIT Biotechnology and Food Research, Espoo, Finland.
Takkinen
K
K
Sizmann
D
D
Söderlund
H
H
Teeri
T T
TT
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000915:Antibody Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006241:Haptens / Q000737:chemistry; D006868:Hydrolysis; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D010081:Oxazolone / Q000031:analogs & derivatives* / Q000276:immunology; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins
D000906:Antibodies; D006241:Haptens; D007128:Immunoglobulin Fragments; D010446:Peptide Fragments; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; C018361:2-phenyloxazolone; D010081:Oxazolone