Comparison of techniques to screen and characterize bacteria-specific hybridomas for high-quality monoclonal antibodies selection

Charlermroj, Ratthaphol; Oplatowska, Michalina; Kumpoosiri, Mallika; Himananto, Orawan; Gajanandana, Oraprapai; Elliott, Christopher T.; Karoonuthaisiri, Nitsara

doi:10.1016/j.ab.2011.10.005

Cited by 16 publications

(11 citation statements)

References 26 publications

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“…For instance, skimmed milk was the best blocking reagent whereas BSA resulted in a high background for a foodborne pathogen antibody array [27]. On the other hand, both skimmed milk and BSA were found to be the most effective blockers for an antibody for hybridoma screening [28]. In addition, several commercially available blocking buffers were evaluated and shown to be effective in eliminating non-specific binding in a microsphere immunoassay [29].…”

Section: Resultsmentioning

confidence: 99%

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

et al. 2013

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Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.

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Section: Resultsmentioning

confidence: 99%

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

et al. 2013

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“…The authors believe incorporation of such high throughput capabilities for screening of phage clones, for both their DNA sequencing and binding capacities, would greatly increase the success rate of obtaining phage clone(s) with the desired target specificity. The use of a high throughout technique such as a microarray system could lead to better and faster screening, as has been demonstrated recently for the high throughput screening of hybridmonas [37]. …”

Section: Discussionmentioning

confidence: 99%

Phage Display-Derived Binders Able to Distinguish Listeria monocytogenes from Other Listeria Species

et al. 2013

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The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens ( Salmonella spp., Escherichia coli and Campylobacter jejuni). Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.

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“…() used an electrochemical detection of specific DNA, combined with the evaluation of respiratory activity, to assess Escherichia coli . Other rapid methods include MALDI‐TOF‐MS (Nicolau et al ., ), hyperspectral scattering (Tao et al ., ), ultrasound and ELISA (Charlermroj et al ., ). These methods show a high level of specificity, but a limitation could be the high cost of the equipment required for the analysis.…”

Section: Introductionmentioning

confidence: 97%

“…Although conventional culture methods are still commonly used to ensure the microbiological quality of milk, some studies were performed to identify culturable microbial communities by means of molecular identification tools (Hantsis-Zacharov & Halpern, 2007); for example, Zhu et al (2012) proposed a combination of impedance and rRNA-based lateral flow assay to assess the contamination by Cronobacter sakazakii in 29 h, whilst Yamanaka et al (2012) used an electrochemical detection of specific DNA, combined with the evaluation of respiratory activity, to assess Escherichia coli. Other rapid methods include MALDI-TOF-MS (Nicolau et al, 2012), hyperspectral scattering (Tao et al, 2012), ultrasound and ELISA (Charlermroj et al, 2012). These methods show a high level of specificity, but a limitation could be the high cost of the equipment required for the analysis.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Pseudomonas spp. through O₂ and CO₂ headspace analysis

Bevilacqua

Corbo

Martino

et al. 2013

Int J of Food Sci Tech

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This paper proposes an approach to determine the level of Pseudomonas spp. in milk, based on the evaluation of the content of oxygen and carbon dioxide produced in the head-space of sealed vials; the research was divided into two phases: model building and preliminary validation. Three different strains of Pseudomonas spp., Ps. putida (wild strain) and Ps. fluorescens (wild and a collection isolates), were used as targets.\ud Data of CO2 and O2 were modeled through a modified positive (CO2) or a negative Gompertz equation (O2), to estimate the Minimum Detection Time (MDT), defined as the time to attain 3% of CO2 (MDT1) or a decrease of the content of O2 by 3% (MDT2). Then, MDT1 and MDT2 were submitted to a linear regression procedure, using cell concentration as independent variable; the correlations "MDT1/cell concentration" and "MDT2/cell concentration" showed high determination coefficients (>0.983). Moreover, the regression procedure pointed out that both MDT1 and MDT2 decreased by ca. 3 h for an increase of cell count of 1 log cfu mL-1. Preliminary validation in milk pointed out that the error associated with the regression line "MDT2/cell concentration" was below 5%

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Comparison of techniques to screen and characterize bacteria-specific hybridomas for high-quality monoclonal antibodies selection

Cited by 16 publications

References 26 publications

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

Phage Display-Derived Binders Able to Distinguish Listeria monocytogenes from Other Listeria Species

Evaluation of Pseudomonas spp. through O₂ and CO₂ headspace analysis

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Comparison of techniques to screen and characterize bacteria-specific hybridomas for high-quality monoclonal antibodies selection

Cited by 16 publications

References 26 publications

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology

Phage Display-Derived Binders Able to Distinguish Listeria monocytogenes from Other Listeria Species

Evaluation of Pseudomonas spp. through O2 and CO2 headspace analysis

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Evaluation of Pseudomonas spp. through O₂ and CO₂ headspace analysis