Development of Specific Recombinant Monoclonal Antibodies Against the Lipopolysaccharide of <i>Ralstonia solanacearum</i> Race 3

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

doi:10.1094/phyto.1998.88.8.795

Cited by 51 publications

(31 citation statements)

References 22 publications

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“…These enriched scFv antibodies were confirmed by polyclonal phage ELISA in which the fourth panning showed higher value than previous panning process even when same concentration of phages (1x10 12 /well) were used. Similar result was reported by Griep, Van Twisk, Beckhoven et al, (1998) for selected scFv antibody against lipopolysaccharide of Ralstonia solanacearum Race-3. The phages obtained from IV panning were used to generate the monoclones.…”

Section: Sensitivity Of Diagnostic Kitsupporting

confidence: 88%

Production of Diagnostic Kit to Detect Cry 2B Antigen by Use of scFv Monoclonal Antibody

Kumar¹,

Moger²,

Rabinal³

et al. 2017

Int.J.Curr.Microbiol.App.Sci

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Section: Sensitivity Of Diagnostic Kitsupporting

confidence: 88%

Production of Diagnostic Kit to Detect Cry 2B Antigen by Use of scFv Monoclonal Antibody

Kumar¹,

Moger²,

Rabinal³

et al. 2017

Int.J.Curr.Microbiol.App.Sci

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“…In most cases, the fragments consist of single-chain variable fragments (scFvs), which are fusions of the variable domains of antibody heavy and light chains through a flexible synthetic linker. Recombinant scFvs have mainly been applied in the medical research field, but have also been described for a number of plant viruses (reviewed in Ziegler and Torrance 2002) and a few other plant pathogens (Griep et al 1998). However, a large scale transition of plant pathogen detection from classical antisera to recombinant antibodies (such as scFvs) has not been made.…”

Section: Introductionmentioning

confidence: 99%

Stable recombinant alpaca antibodies for detection of Tulip virus X

Beekwilder¹,

Houwelingen²,

Beckhoven

et al. 2008

Eur J Plant Pathol

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For detection of the plant pathogenic Tulip virus X (TuVX), a panel of six recombinant antibodies was identified. To this end, a repertoire of variable domains from heavy-chain immunoglobulins (VHH) was cloned from an alpaca, which had been immunized with TuVX. Binding domains were selected by phage display and panning on immobilized TuVX particles. Recombinant VHH antibodies were tested for sensitivity in a sandwich ELISA, and were demonstrated to be readily able to distinguish TuVX-infected tulip leaf material from uninfected leafs. No cross-reactivity of the VHH antibodies to related flexiviridae was observed. Recombinant VHHs maintained their reactivity upon storage at −20°C for over a year. The effect of incubation at higher temperatures for prolonged time was studied. Two out of three VHH proteins retained activity after several weeks of storage at 37°C.

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“…• C. A very similar approach to our work using antibody phage-display was done by Griep et al who used purified LPS of Ralstonia solanacearum Race 3 for antibody selection [18]. The selected antibodies were used in combination with a polyclonal capture antisera in order to detect bacterial cells by sandwich-ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…High sensitivity of 5 × 10 3 cfu/ml was reached, but a polyclonal antiserum was still needed, adding the disadvantages of limited resource and complex undefined composition. Furthermore, this LPS antibody crossreacted with other organisms, rendering very much its relevance for diagnostics [18].…”

Section: Discussionmentioning

confidence: 99%

Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity

Kühn

Thiem

Steinert

et al. 2019

HAB

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Abstract. Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a lifethreatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella. The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae, Pseudomonadaceae or Clostridiaceae. Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring.

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Development of Specific Recombinant Monoclonal Antibodies Against the Lipopolysaccharide of Ralstonia solanacearum Race 3

Cited by 51 publications

References 22 publications

Production of Diagnostic Kit to Detect Cry 2B Antigen by Use of scFv Monoclonal Antibody

Production of Diagnostic Kit to Detect Cry 2B Antigen by Use of scFv Monoclonal Antibody

Stable recombinant alpaca antibodies for detection of Tulip virus X

Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity

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