Optimization and application of a dithiobis-succinimidyl propionate-modified immunosensor platform to detect Listeria monocytogenes in chicken skin

Park, Mi-Kyung; Park, Jang Won; Oh, Jung Hyun

doi:10.1016/j.snb.2012.04.017

Cited by 19 publications

(17 citation statements)

References 26 publications

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“…The indirect ELISA method involved the primary antibodies without enzyme and the secondary antibodies (which reacted with primary antibodies) combined with enzymes. When the enzymes react with substrates that can produce a fluorescent compound, color reaction occurs between the substrates and the enzymes combined with secondary antibodies [18]. The concentration of the tested bacterial species was approximately 1 × 10 8 CFU/mL.…”

Section: Discussionmentioning

confidence: 99%

“…The specific objectives of this research are to (1) investigate the binding specificity between the target Staphylococcus aureus and polyclonal anti-Staphylococcus aureus antibodies for biosensor applications; (2) develop a fabrication process for SWCNT-based biosensor; and (3) characterize the properties of SWCNT-based biosensor to detect Staphylococcus aureus. The specificity of pAbs was compared with commercially available anti-staphylococcus aureus using an indirect ELISA following the procedures described in the previous study [18]. S. aureus was cultivated in nutrient broth (Difco Laboratories, Detroit, MI, USA), and the other bacteria were cultivated in tryptic soy broth (Difco Laboratories, Detroit, MI, USA) in a shaking incubator at 37 ∘ C for 16 h. The grown and collected bacteria were washed three times with 10 mM PBS buffer (pH 7.4) by centrifugation at 5,000 rpm for 1 min at 4 ∘ C. An aliquot of 100 L anti-Staphylococcus aureus antibodies diluted ten thousandfold with PBS buffer was taken and incubated in each well of the microplate (SPL, Pocheon, South Korea).…”

Section: Introductionmentioning

confidence: 99%

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Development of Single-Walled Carbon Nanotube-Based Biosensor for the Detection of Staphylococcus aureus

Choi

Lee

Park

et al. 2017

Journal of Food Quality

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The goal of this research is to develop a single-walled carbon nanotube-(SWCNT-) based biosensor to detect Staphylococcus aureus. The specificity of 11 bacteria and polyclonal anti-Staphylococcus aureus antibodies (pAbs) was determined using an indirect ELISA. The pAbs were immobilized onto sensor platform after the hybridization of 1-pyrenebutanoic acid succinimidyl ester (PBASE). The resistance difference (Δ ) was calculated using a potentiostat. The bacteria detected by the biosensor were observed using a scanning electron microscope (SEM). The optimum concentration of SWCNTs on the platform was determined to be 0.1 mg/mL. The binding of pAbs with S. aureus resulted in a significant increase in resistance value of the biosensor ( < 0.05). The SEM images confirmed the specific binding of S. aureus on the biosensor. The SWCNT-based biosensor was able to detect S. aureus with a limit of detection (LOD) of 4 log CFU/mL.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of Single-Walled Carbon Nanotube-Based Biosensor for the Detection of Staphylococcus aureus

Choi

Lee

Park

et al. 2017

Journal of Food Quality

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“…11 In this research, L. monocytogenes were detected after a 10-h enrichment period by the DSP-modified immunosensor alone ( Figure 4). However, when IMS was coupled with the DSP-modified immunosensor, the incubation time decreased to 7.5 h, although no significant difference was noted in the number of detected L. monocytogenes.…”

Section: Optimization Of Ims-mentioning

confidence: 59%

“…11 The conjugation of MBs to prepare immunomagnetic beads (IMBs) was performed by immobilizing MBs (Bioclone Inc., San Diego, CA, USA) covalently with anti-Listeria pAbs. According to the manufacturer's instructions, MBs were reacted with coupling buffer (10 mM potassium phosphate, 0,15 M NaCl, pH 5.5) and coupled with Anti-Listeria pAbs in the presence of EDC [1-ethyl-3 (3-dimethylaminopropyl) carbodiimide] as a coupling agent.…”

Section: Bacterialmentioning

confidence: 99%

“…DSP modification of the immunosensor greatly improved the efficacy of the immunosensor by decreasing the possibility of random orientation of antibodies and maintaining the conformational stability of the antibody on the immunosensor platform when applied to Escherichia coli O157:H7 in turnip greens 9 and L. monocytogenes in chicken products. 11 However, when applied to real food systems, the sensitivity and reliability of DSP-modified immunosensors greatly depend on impurities of the food matrix detected by the biosensor. Ubiquitous food particles in food matrix such as lipids or other components that are present z E-mail: junhyunoh@smu.ac.kr in chicken products interfere with the specific binding between target bacteria and antibodies immobilized on the immunosensor platform.…”

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confidence: 99%

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Immunomagnetic Bead Separation Coupled with a Dithiobis-Succinimidyl Propionate (DSP)-Modified Immunosensor to DetectListeria monocytogenesin Chicken Skin

Park

2014

J. Electrochem. Soc.

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This study aimed to determine the optimal conditions for coupling immunomagnetic bead separation (IMS) with a dithiobissuccinimidyl propionate (DSP)-modified immunosensor and apply the IMS-coupled DSP-modified immunosensor to detect Listeria monocytogenes in chicken skin. Specific anti-Listeria polyclonal antibodies (pAbs) against three L. monocytogenes species were covalently immobilized on magnetic beads (MBs) to prepare immunomagnetic beads (IMBs). The optimal pAb concentration, incubation time, buffer type and pH, amount of MBs, and elution buffer for IMS were determined to be 1.0 mg/mL, 30 min, PBS buffer with a pH range from 6.5 to 7.5, 30 μL, and citric acid (pH 3.0), respectively. The number of L. monocytogenes after IMS application was 6.0 log colony-forming unit (CFU)/g during the 15-h enrichment period, as compared to 7.1 log CFU/g without IMS application. IMS coupled with a DSP-modified immunosensor decreased the incubation time to 7.5 h with a limit of detection of 10 3 CFU/25 g chicken and improved the sensitivity of the immunosensor, exhibiting a correlation coefficient (R 2 ) of 0.95. Microscopic images of the immunosensor coupled with IMS revealed less binding of food particles on MBs than binding without IMS treatment.

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Probing antibody surface density and analyte antigen incubation time as dominant parameters influencing the antibody-antigen recognition events of a non-faradaic and diffusion-restricted electrochemical immunosensor

et al. 2020

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Electrochemical sensors based on antibody-antigen recognition events are commonly used for the rapid, label-free, and sensitive detection of various analytes. However, various parameters at the bioelectronic interface, i.e., before and after the probe (such as an antibody) assembly onto the electrode, have a dominant influence on the underlying detection performance of analytes (such as an antigen). In this work, we thoroughly investigate the dependence of the bioelectronic interface characteristics on parameters that have not been investigated in depth: the antibody density on the electrode's surface and the antigen incubation time. For this important aim, we utilized the sensitive non-faradaic electrochemical impedance spectroscopy method. We showed that as the incubation time of the antigen-containing drop solution increased, a decrease was observed in both the solution resistance and the diffusional resistance with reflecting boundary elements, as well as the capacitive magnitude of a constant phase element, which decreased at a rate of 160 ± 30 kΩ/min, 800 ± 100 mΩ/min, and 520 ± 80 pF × s (α-1) /min, respectively. Using atomic force microscopy, we also showed that high antibody density led to thicker electrode coating than low antibody density, with rootmean-square roughness values of 2.2 ± 0.2 nm versus 1.28 ± 0.04 nm, respectively. Furthermore, we showed that as the antigen accumulated onto the electrode, the solution resistance increased for high antibody density and decreased for low antibody density. Finally, the antigen detection performance test yielded a better limit of detection for low antibody density than for high antibody density (0.26 μM vs 2.2 μM). Overall, we show here the importance of these two factors and how changing one parameter can drastically affect the desired outcome.

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Optimization and application of a dithiobis-succinimidyl propionate-modified immunosensor platform to detect Listeria monocytogenes in chicken skin

Cited by 19 publications

References 26 publications

Development of Single-Walled Carbon Nanotube-Based Biosensor for the Detection of Staphylococcus aureus

Development of Single-Walled Carbon Nanotube-Based Biosensor for the Detection of Staphylococcus aureus

Immunomagnetic Bead Separation Coupled with a Dithiobis-Succinimidyl Propionate (DSP)-Modified Immunosensor to DetectListeria monocytogenesin Chicken Skin

Probing antibody surface density and analyte antigen incubation time as dominant parameters influencing the antibody-antigen recognition events of a non-faradaic and diffusion-restricted electrochemical immunosensor

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