Experimental section

O’Kennedy, Richard; Byrne, M.; Ó’Fágáin, Ciarán; Berns, Gerard

doi:10.1016/0307-4412(90)90219-e

Cited by 15 publications

(10 citation statements)

References 2 publications

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“…However, a vigorous washing regimen has often been reported to cause partial desorption of passively adsorbed antigen/antibody from the wells, thereby affecting the sensitivity, limit-of-detection, and reproducibility of the assay. 3,6 The propensity of desorption is more pronounced in competitive ELISA, especially when a small molecule is adsorbed onto the well plate. Unlike high molecular weight antibodies, small molecules fail to establish a binding interaction with PS well and are washed away easily.…”

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confidence: 99%

Plate-Adherent Nanosubstrate for Improved ELISA of Small Molecules: A Proof of Concept Study

et al. 2020

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Enzyme-linked immunosorbent assay (ELISA) is a widely used technique for detecting and quantifying target analytes in clinical and research laboratories. One of the main drawbacks of ELISA is the involvement of multiple washing steps that desorbs the capture antigen/antibody off the polystyrene plate, thereby producing inconsistent and erroneous data. To overcome the problem of desorption, we hypothesized that gelatin nanoparticles (GelNP) could serve as a "plate-adherent" substrate to irreversibly adhere the capture antigen/antibody of interest. We tested our hypothesis using GelNP-based substrate (Gel−BSA− OHG) to adhere 8-hydroxy-2′-deoxyguanosine (8-OHdG) to the polystyrene plate and assayed this molecule using the ELISA technique. The stability and ELISA performance of Gel−BSA−OHG was evaluated in comparison to the conventional substrate (BSA−OHG). Importantly, the Gel−BSA−OHG substrate was found to be more wash-resistant and consequently resulted in improved sensitivity, accuracy, and precision in the ELISA analysis of 8-OHdG. Finally, the scope of Gel−BSA−OHG substratebased ELISA for clinical application was demonstrated by validating its ability to detect 8-OHdG in an artificial urine sample with high specificity.

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confidence: 99%

Plate-Adherent Nanosubstrate for Improved ELISA of Small Molecules: A Proof of Concept Study

et al. 2020

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“…In an attempt to reduce costs without compromising on the detection sensitivity, [67][68][69] Compared to HPLC, immunoassays are cheaper, faster and can be easily performed using commercial kits and automated analyzers (e.g. TDx from Abbott Laboratories for FPIA) with good precision, accuracy and sensitivity; therefore it is widely adopted in most routine laboratories.…”

Section: Hplcmentioning

confidence: 99%

“…There are two types of EIA, namely EMIT and enzyme-linked immunosorbent assay (ELISA), which are classified as homogeneous and heterogeneous assays, respectively. 69,78 The latter involves a physical separation step whereby bound complexes formed after incubation are retained on a solid phase and unbound materials are washed away. 78 This is not necessary in the homogeneous assay which greatly enhances its efficiency.…”

Section: Eiamentioning

confidence: 99%

“…78 This is not necessary in the homogeneous assay which greatly enhances its efficiency. 67,69 Similar to FPIA, commercial kits for EMIT are available and the operation can be automated (e.g. Du Pont's aca ® system, Syva's Solaris ® system and Cobas Bio™ centrifugal analyser from Roche Analytical Instruments, Inc.).…”

Section: Eiamentioning

confidence: 99%

“…to assay conditions, shelf life, storage and transport requirements) and batch-to-batch consistency, which are often cited as inadequacies of immunoassays. 69,84 To resolve this, molecularly imprinted materials (MIM) have been developed as synthetic alternatives to natural antibodies.…”

Section: Eiamentioning

confidence: 99%

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Design and development of novel fluorescence-based detection methods for bio-analytes

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I would like to express my gratitude to many people who have given me their help and support in one way or another throughout my four years of research studies. First of all, I would like to thank Nanyang Technological University for the research scholarship and great opportunity to pursue my graduate studies in such a wonderful environment. I would like to thank the Division of Chemistry and Biological Chemistry (CBC) for the excellent research infrastructure and the devoted staff for their kind assistance that has helped my research experience to be a lot smoother. I would like to express my heartfelt thanks to my supervisor, Assoc. Prof. Edwin Kok Lee Yeow. I know that sometimes you find it difficult to communicate your point across to me, but thank you for not giving up and still being so patient to teach and guide me. You taught me how to be more open-minded to consider different questions and not to reject it hastily. You also taught me how to look at the bigger picture whenever I am stuck with the details of my work. Thank you for guiding me through the different projects and the whole process of publishing a paper with your expertise and hard work. I would also like to thank my co-supervisor, Assoc. Prof. Bengang Xing, for your encouragement and advice throughout the four years. I am very grateful to my mentor and colleague, Dr. Xiangyang Wu. You are a very experienced and knowledgeable researcher. Thank you for teaching me many things from my first year of PhD and always guiding me and helping me to solve many problems. You have a lot of ideas and you are persistent to work on those ideas. The way you conduct research taught me how a research life is and should be like. I would like to thank my collaborators, Assoc. Prof. Howe Siang Tan, Mr.

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