Analysis of direct immobilized recombinant protein G on a gold surface

Kim, Hyunhee; Kang, Da-Yeon; Goh, Hyun-Jeong; Oh, Byung-Keun; Singh, Ravindra Pratap; Oh, Soo-Min; Choi, Jeong‐Woo

doi:10.1016/j.ultramic.2008.04.049

Cited by 16 publications

(14 citation statements)

References 16 publications

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“…For the detection of AFB1, we first installed a pre-cleaned Au chip in the palm-sized SPR system, and the running buffer (PBS) and 0.1 mg/mL cysteine-protein G were flowed through both the test and control channel. Cysteine-protein G can bind to the Au chip through Au–S bonding [ 27 , 28 ]. Second, 0.1 mg/mL polyclonal anti-AFB1 antibody was immobilized on the Au chip through the binding of protein G to the Fc region of the antibody.…”

Section: Resultsmentioning

confidence: 99%

On-Site Detection of Aflatoxin B1 in Grains by a Palm-Sized Surface Plasmon Resonance Sensor

Moon

Byun

Kim

et al. 2018

Sensors

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Aflatoxins (AFs) are highly toxic compounds that can cause both acute and chronic toxicity in humans. Aflatoxin B1 (AFB1) is considered the most toxic of AFs. Therefore, the rapid and on-site detection of AFB1 is critical for food safety management. Here, we report the on-site detection of AFB1 in grains by a portable surface plasmon resonance (SPR) sensor. For the detection of AFB1, the surface of an SPR Au chip was sequentially modified by cysteine-protein G, AFB1 antibody, and bovine serum albumin (BSA). Then, the sample solution and AFB1-BSA conjugate were flowed onto the Au chip in serial order. In the absence of AFB1, the SPR response greatly increased due to the binding of AFB1-BSA on the Au chip. In the presence of AFB1, the SPR response showed little change because the small AFB1 molecule binds on the Au chip instead of the large AFB1-BSA molecule. By using this portable SPR-based competitive immunoassay, the sensor showed low limits of detection (2.51 ppb) and quantification (16.32 ppb). Furthermore, we successfully detected AFB1 in rice, peanut, and almond samples, which suggests that the proposed sensing method can potentially be applied to the on-site monitoring of mycotoxins in food.

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Section: Resultsmentioning

confidence: 99%

On-Site Detection of Aflatoxin B1 in Grains by a Palm-Sized Surface Plasmon Resonance Sensor

Moon

Byun

Kim

et al. 2018

Sensors

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“…3.02 AE 1.26 nm and 3.42 AE 1.62 nm, respectively. 50 Highly ordered and dense packed SAM have fewer defects and are better stabilized by van der Waals forces. 9.8 AE 2.8 nm and 9.76 AE 2.25 nm as compared to that of TGA and 3MPA.…”

Section: Analysis Of Surface Thickness Measurement Aer Antibody-antimentioning

confidence: 99%

Studies on varying n-alkanethiol chain lengths on a gold coated surface and their effect on antibody–antigen binding efficiency

et al. 2015

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Self-assembled monolayer (SAM) of n-alkanethiols of different chain lengths (n ¼ 2, 3, 6, 11, 16) on a gold surface are used to immobilize antibodies which in turn bind to a antigen. The antibody and antigen used in this study have similar molecular weights i.e. $150 kDa. The antibody [1.5 mg cm À2 ] immobilized varied with the surface packing density of the SAM of carboxylic acid-terminated n-alkanethiols of different lengths. In comparison, the efficiency of antibody immobilization was lowest on the loosely packed SAM of n-alkanethiols (n # 3) and the highest on the densely packed SAM of n-alkanethiols (n $ 11). However, increased immobilization of antibodies with increasing chain length of the n-alkanethiols [n > 11], did not result in a corresponding increase in antigen binding. An attempt to explain this phenomenon based on packing density and an orientation of the captured antibody is presented.www.rsc.org/advances 80480 | RSC Adv., 2015, 5, 80480-80487This journal is

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“…Protein A is a recombinant protein that can bind strongly to the Fc region of an antibody, and therefore, a well-oriented antibody configuration can be established through their affinity binding 37 , 38 . Here, the protein A was immobilized on the surface of the bare CBC and AuNP-CBC through EDC/Sulfo-NHS chemistry by covalent binding between amino groups of the protein A and the activated carboxyl groups.…”

Section: Resultsmentioning

confidence: 99%

Toward a nanopaper-based and solid phase immunoassay using FRET for the rapid detection of bacteria

Heli

Ajji

2020

Sci Rep

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in this study, we propose a novel sensitive solid-based immunosensor developed on a plasmonic nanopaper platform for the detection of Escherichia coli (E. coli) bacteria. this plasmonic nanopaper that comprises of carboxylated bacterial cellulose (cBc) impregnated with gold nanoparticles (Aunp-cBc), employed as a quencher and a sustainable functionalized platform to be conjugated with protein A. thus, the conjugated protein A allows the aligned linkage of eAb-QD (anti-E. coli conjugated quantum dot) and eAb-Af (anti-E. coli conjugated Alexa Fluor 488). Interestingly, once E. coli was captured by the Aunp-cBc/eAb-QD or Aunp-cBc/eAb-Af, the energy transfer from the QD or Alexa Fluor fluorophores is triggered due to the conformational change in the antibody structure and this, in turn, causes a decrease in the distance between fluorophores and the quencher nanopaper and, therefore diminishing their photoluminescence. the immunosensors performed successfully to recognize E. coli at concentrations as low as 50 CFU mL −1 in the standard buffer. The examined functionality of the immunosensors in a real matrix such as chicken extract and lettuce juice demonstrated a highly efficient response while QD is the main fluorophore with a limit of detection around 100 CFU mL −1. During the last decades, the outbreak of foodborne diseases has seriously threatened people lives worldwide and led to a considerable number of hospitalizations and even deaths 1. Indeed, the incidence of foodborne infection is a result of ingesting food contaminated by foodborne pathogens such as bacteria, viruses, and parasites. To prevent its subsequence, precise monitoring and early recognition of pathogen would be the primary step to be ensured about the safety of our food 2. Upon comparing various categories of biosensors developed for pathogen detection, immunosensors have attracted plenty of attention due to their flexibility and compatibility in numerous transducer techniques such as electrochemical (voltammetry, impedancemetry, potentiometry), optical (fluorescence, SPR) and mass-based 3-5. The excellence of an immunosensor is directly dominated by the affinity binding of the antigen-antibody that this makes it uniquely sensitive and selective toward a specific target 6. Being fast, selective and sensitive, low-priced, in-situ and user-friendly, immunosensors can fully address the challenges associated with traditional bacteria detection approaches, such as; costly analysis, long processing time, and needs in scientific expertise 7. Given these benefits, its applications have been widely explored in the environment, medical and, food industries that designed either on a solid-based platform or solution-phase method 8,9. Assuredly, the integration of nanomaterials with immunosensing techniques has caused a huge improvement in its functionality and sensitivity 10-12. Nanoscale materials, with a size less than 100 nm are endowed with unique physical and chemical properties such as; high surface area, optical, and electrical characteristics, which ...

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Analysis of direct immobilized recombinant protein G on a gold surface

Cited by 16 publications

References 16 publications

On-Site Detection of Aflatoxin B1 in Grains by a Palm-Sized Surface Plasmon Resonance Sensor

On-Site Detection of Aflatoxin B1 in Grains by a Palm-Sized Surface Plasmon Resonance Sensor

Studies on varying n-alkanethiol chain lengths on a gold coated surface and their effect on antibody–antigen binding efficiency

Toward a nanopaper-based and solid phase immunoassay using FRET for the rapid detection of bacteria

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