Properties of a single-chain antibody containing different linker peptides

Kaija Alfthan (Corresponding Author), Kristiina Takkinen, Dorothea Sizmann, Hans Söderlund, Tuula Teeri

Research output: Contribution to journalArticleScientificpeer-review

59 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)725-731
JournalProtein Engineering
Volume8
Issue number7
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

Fingerprint

Single-Chain Antibodies
Antibodies
Peptides
Immunoglobulin Variable Region
Immunoglobulin Fragments
Haptens
Amino acids
Amino Acids
Cellulase
Hinges
Joining
Peptide Hydrolases
Escherichia coli
Enzyme-Linked Immunosorbent Assay
Derivatives

Cite this

Alfthan, Kaija ; Takkinen, Kristiina ; Sizmann, Dorothea ; Söderlund, Hans ; Teeri, Tuula. / Properties of a single-chain antibody containing different linker peptides. In: Protein Engineering. 1995 ; Vol. 8, No. 7. pp. 725-731.
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abstract = "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.",
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Properties of a single-chain antibody containing different linker peptides. / Alfthan, Kaija (Corresponding Author); Takkinen, Kristiina; Sizmann, Dorothea; Söderlund, Hans; Teeri, Tuula.

In: Protein Engineering, Vol. 8, No. 7, 1995, p. 725-731.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Properties of a single-chain antibody containing different linker peptides

AU - Alfthan, Kaija

AU - Takkinen, Kristiina

AU - Sizmann, Dorothea

AU - Söderlund, Hans

AU - Teeri, Tuula

N1 - Project code: BEL2026

PY - 1995

Y1 - 1995

N2 - 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.

AB - 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.

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