Development of specific recombinant monoclonal antibodies against the lipopolysaccharides of Ralstonia solanacearum race 3

R.A. Griep; C. van Twisk; J.R.C.M. van Beckhoven; J.M. van der Wolf; A. Schots

doi:10.1094/PHYTO.1998.88.8.795

Development of specific recombinant monoclonal antibodies against the lipopolysaccharides of Ralstonia solanacearum race 3

R.A. Griep, C. van Twisk, J.R.C.M. van Beckhoven, J.M. van der Wolf, A. Schots

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 × 103 bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.

Original language	English
Pages (from-to)	795-803
Journal	Phytopathology
Volume	88
Issue number	8
DOIs	https://doi.org/10.1094/PHYTO.1998.88.8.795
Publication status	Published - 1998

Keywords

Bacterial wilt
Burkholderia solanacearum
Pseudomonas solanacearum

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@article{887f2e9ec17d4baca394203a3d95d34f,

title = "Development of specific recombinant monoclonal antibodies against the lipopolysaccharides of Ralstonia solanacearum race 3",

abstract = "Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 × 103 bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.",

keywords = "Bacterial wilt, Burkholderia solanacearum, Pseudomonas solanacearum",

author = "R.A. Griep and {van Twisk}, C. and {van Beckhoven}, J.R.C.M. and {van der Wolf}, J.M. and A. Schots",

year = "1998",

doi = "10.1094/PHYTO.1998.88.8.795",

language = "English",

volume = "88",

pages = "795--803",

journal = "Phytopathology",

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T1 - Development of specific recombinant monoclonal antibodies against the lipopolysaccharides of Ralstonia solanacearum race 3

AU - Griep, R.A.

AU - van Twisk, C.

AU - van Beckhoven, J.R.C.M.

AU - van der Wolf, J.M.

AU - Schots, A.

PY - 1998

Y1 - 1998

N2 - Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 × 103 bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.

AB - Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 × 103 bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.

KW - Bacterial wilt

KW - Burkholderia solanacearum

KW - Pseudomonas solanacearum

U2 - 10.1094/PHYTO.1998.88.8.795

DO - 10.1094/PHYTO.1998.88.8.795

M3 - Article

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EP - 803

JO - Phytopathology

JF - Phytopathology

IS - 8

ER -

Development of specific recombinant monoclonal antibodies against the lipopolysaccharides of Ralstonia solanacearum race 3

Abstract

Keywords

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