Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library

Tuomas Heiskanen, Xiao Dong Li, Jussi Hepojoki, Elisabeth Gustafsson, Ake Lundkvist, Antti Vaheri, Hilkka Lankinen

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have previously selected a peptide insert FPCDRLSGYWERGIPSPCVR recognizing the Puumala virus (PUUV) G2-glycoprotein-specific neutralizing monoclonal antibody (MAb) 1C9 with Kd of 2.85 × 10-8 from a random peptide library X2CX14CX2 expressed on the pIII protein of the filamentous phage fd-tet. We have now created a second-generation phage-displayed peptide library in which each amino acid of the peptide was mutated randomly to another with a certain probability. Peptides were selected for higher affinity for MAb 1C9 and for a common binding motif for MAb 4G2 having an overlapping epitope with MAb 1C9 in G2 glycoprotein. The resulting peptides were synthesized as spots on cellulose membrane. Amino acid changes which improved the reactivity of the peptides to MAb 1C9 were combined in the peptide ATCDKLFGYYERGIPLPCAL with Kd of 1.49 × 10-9 in biosensor measurements. Our results show that the binding properties of peptides, the affinity and the specificity can be improved and the binding specificity determining amino acids and structural factors can be analyzed by combining binding assays with synthetic peptides on membrane with the use of second-generation phage display libraries.

Original languageEnglish (US)
Pages (from-to)443-450
Number of pages8
JournalProtein Engineering
Volume16
Issue number6
StatePublished - Jun 1 2003

Fingerprint

Puumala virus
Bacteriophages
Viruses
Peptides
Monoclonal antibodies
Monoclonal Antibodies
Peptide Library
Amino acids
Glycoproteins
Amino Acids
Bacteriophage M13
Membranes
Biosensing Techniques
Epitopes
Neutralizing Antibodies
Biosensors
Cellulose
Libraries
Assays
Display devices

Keywords

  • Epitope
  • Hantavirus
  • Peptide
  • Phage
  • Second-generation library

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

Cite this

Heiskanen, T., Li, X. D., Hepojoki, J., Gustafsson, E., Lundkvist, A., Vaheri, A., & Lankinen, H. (2003). Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library. Protein Engineering, 16(6), 443-450.

Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library. / Heiskanen, Tuomas; Li, Xiao Dong; Hepojoki, Jussi; Gustafsson, Elisabeth; Lundkvist, Ake; Vaheri, Antti; Lankinen, Hilkka.

In: Protein Engineering, Vol. 16, No. 6, 01.06.2003, p. 443-450.

Research output: Contribution to journalArticle

Heiskanen, T, Li, XD, Hepojoki, J, Gustafsson, E, Lundkvist, A, Vaheri, A & Lankinen, H 2003, 'Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library', Protein Engineering, vol. 16, no. 6, pp. 443-450.
Heiskanen T, Li XD, Hepojoki J, Gustafsson E, Lundkvist A, Vaheri A et al. Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library. Protein Engineering. 2003 Jun 1;16(6):443-450.
Heiskanen, Tuomas ; Li, Xiao Dong ; Hepojoki, Jussi ; Gustafsson, Elisabeth ; Lundkvist, Ake ; Vaheri, Antti ; Lankinen, Hilkka. / Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library. In: Protein Engineering. 2003 ; Vol. 16, No. 6. pp. 443-450.
@article{eee0bf4f4567436aa15e6de4b965f778,
title = "Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library",
abstract = "We have previously selected a peptide insert FPCDRLSGYWERGIPSPCVR recognizing the Puumala virus (PUUV) G2-glycoprotein-specific neutralizing monoclonal antibody (MAb) 1C9 with Kd of 2.85 × 10-8 from a random peptide library X2CX14CX2 expressed on the pIII protein of the filamentous phage fd-tet. We have now created a second-generation phage-displayed peptide library in which each amino acid of the peptide was mutated randomly to another with a certain probability. Peptides were selected for higher affinity for MAb 1C9 and for a common binding motif for MAb 4G2 having an overlapping epitope with MAb 1C9 in G2 glycoprotein. The resulting peptides were synthesized as spots on cellulose membrane. Amino acid changes which improved the reactivity of the peptides to MAb 1C9 were combined in the peptide ATCDKLFGYYERGIPLPCAL with Kd of 1.49 × 10-9 in biosensor measurements. Our results show that the binding properties of peptides, the affinity and the specificity can be improved and the binding specificity determining amino acids and structural factors can be analyzed by combining binding assays with synthetic peptides on membrane with the use of second-generation phage display libraries.",
keywords = "Epitope, Hantavirus, Peptide, Phage, Second-generation library",
author = "Tuomas Heiskanen and Li, {Xiao Dong} and Jussi Hepojoki and Elisabeth Gustafsson and Ake Lundkvist and Antti Vaheri and Hilkka Lankinen",
year = "2003",
month = "6",
day = "1",
language = "English (US)",
volume = "16",
pages = "443--450",
journal = "Protein Engineering, Design and Selection",
issn = "1741-0126",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - Improvement of binding of Puumala virus neutralization site resembling peptide with a second-generation phage library

AU - Heiskanen, Tuomas

AU - Li, Xiao Dong

AU - Hepojoki, Jussi

AU - Gustafsson, Elisabeth

AU - Lundkvist, Ake

AU - Vaheri, Antti

AU - Lankinen, Hilkka

PY - 2003/6/1

Y1 - 2003/6/1

N2 - We have previously selected a peptide insert FPCDRLSGYWERGIPSPCVR recognizing the Puumala virus (PUUV) G2-glycoprotein-specific neutralizing monoclonal antibody (MAb) 1C9 with Kd of 2.85 × 10-8 from a random peptide library X2CX14CX2 expressed on the pIII protein of the filamentous phage fd-tet. We have now created a second-generation phage-displayed peptide library in which each amino acid of the peptide was mutated randomly to another with a certain probability. Peptides were selected for higher affinity for MAb 1C9 and for a common binding motif for MAb 4G2 having an overlapping epitope with MAb 1C9 in G2 glycoprotein. The resulting peptides were synthesized as spots on cellulose membrane. Amino acid changes which improved the reactivity of the peptides to MAb 1C9 were combined in the peptide ATCDKLFGYYERGIPLPCAL with Kd of 1.49 × 10-9 in biosensor measurements. Our results show that the binding properties of peptides, the affinity and the specificity can be improved and the binding specificity determining amino acids and structural factors can be analyzed by combining binding assays with synthetic peptides on membrane with the use of second-generation phage display libraries.

AB - We have previously selected a peptide insert FPCDRLSGYWERGIPSPCVR recognizing the Puumala virus (PUUV) G2-glycoprotein-specific neutralizing monoclonal antibody (MAb) 1C9 with Kd of 2.85 × 10-8 from a random peptide library X2CX14CX2 expressed on the pIII protein of the filamentous phage fd-tet. We have now created a second-generation phage-displayed peptide library in which each amino acid of the peptide was mutated randomly to another with a certain probability. Peptides were selected for higher affinity for MAb 1C9 and for a common binding motif for MAb 4G2 having an overlapping epitope with MAb 1C9 in G2 glycoprotein. The resulting peptides were synthesized as spots on cellulose membrane. Amino acid changes which improved the reactivity of the peptides to MAb 1C9 were combined in the peptide ATCDKLFGYYERGIPLPCAL with Kd of 1.49 × 10-9 in biosensor measurements. Our results show that the binding properties of peptides, the affinity and the specificity can be improved and the binding specificity determining amino acids and structural factors can be analyzed by combining binding assays with synthetic peptides on membrane with the use of second-generation phage display libraries.

KW - Epitope

KW - Hantavirus

KW - Peptide

KW - Phage

KW - Second-generation library

UR - http://www.scopus.com/inward/record.url?scp=0043268749&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0043268749&partnerID=8YFLogxK

M3 - Article

VL - 16

SP - 443

EP - 450

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

SN - 1741-0126

IS - 6

ER -