Improving immunobinding using oriented immobilization of an oxidized antibody

Kang, Jung-Ha; Choi, Heeseon; Hwang, Sang Youn; Han, Sang Ho; Jeon, Jun Yeoung; Lee, Eun Kyu

doi:10.1016/j.chroma.2007.05.023

Cited by 45 publications

(25 citation statements)

References 25 publications

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“…The F c fraction will be oriented to the bead surface, meanwhile the F ab fraction will be free to recognize spore antigens, and thus, even when antibody concentration on bead surface is low, the better orientation would make easier the binding of spores [44]. The influence of coating method on antibody orientation was more evident when antibody concentration used for bead coating was not optimum, showing aminated beads a slightly better separation capacity than carboxylated beads (with other conditions being analogous), since aminated beads bound antibodies by means of carbohydrate residues of F c region [45,46]. Other authors have also observed this effect, whose results showed that binding capacity of randomly bounded antibodies was twice or three times lower than well-oriented antibodies [47,48].…”

Section: Resultsmentioning

confidence: 99%

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

et al. 2011

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Nowadays, the reference method for the detection of Clostridium tyrobutyricum in milk is the most-probable-number method, a very time-consuming and non-specific method. In this work, the suitability of the use of superparamagnetic beads coated with specific antibodies and peptides for bioseparation and concentration of spores of C. tyrobutyricum has been assessed. Peptide or antibody functionalized nanoparticles were able to specifically bind C. tyrobutyricum spores and concentrate them up to detectable levels. Moreover, several factors, such as particle size (200 nm and 1 μm), particle derivatization (aminated and carboxylated beads), coating method, and type of ligand have been studied in order to establish the most appropriate conditions for spore separation. Results show that concentration of spore is favored by a smaller bead size due to the wider surface of interaction in relation to particle volume. Antibody orientation, related to the binding method, is also critical in spore recovery. However, specific peptides seem to be a better ligand than antibodies, not only due to the higher recovery ratio of spores obtained but also due to the prolonged stability over time, allowing an optimal recovery of spores up to 3 weeks after bead coating. These results demonstrate that specific peptides bound to magnetic nanoparticles can be used instead of traditional antibodies to specifically bind C. tyrobutyricum spores being a potential basis for a rapid method to detect this bacterial target.

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Section: Resultsmentioning

confidence: 99%

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

et al. 2011

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“…The phosphors were then washed 3 times in pure anhydrous ethanol, then dried for 3 hours under reduced pressure and elevated temperature (≈37°C) with the SpeedVac concentrator. Monoclonal mouse anti-β hCG antibodies (Arista Biologicals, Item # ABBCG-0402) were oxidized to introduce aldehydes 49,50 for coupling by first transferring the antibodies into pH 5.4 sodium acetate buffer with a 7k Zeba Spin Desalting Column (ThermoFisher, Catalog # 89882). A 100 mM stock solution of sodium periodate was prepared in sodium acetate buffer, then added to the buffer-exchanged antibodies so that the final sodium periodate concentration was 10 mM with an IgG concentration of ≈1 mg/mL.…”

Section: Methodsmentioning

confidence: 99%

A low-cost smartphone-based platform for highly sensitive point-of-care testing with persistent luminescent phosphors

et al. 2017

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Through their computational power and connectivity, smartphones are poised to rapidly expand telemedicine and transform healthcare by enabling better personal health monitoring and rapid diagnostics. Recently, a variety of platforms have been developed to enable smartphone-based point-of-care testing using imaging-based readout with the smartphone camera as the detector. Fluorescent reporters have been shown to improve the sensitivity of assays over colorimetric labels, but fluorescence readout necessitates incorporating optical hardware into the detection system, adding to the cost and complexity of the device. Here we present a simple, low-cost smartphone-based detection platform for highly sensitive luminescence imaging readout of point-of-care tests run with persistent luminescent phosphors as reporters. The extremely bright and long-lived emission of persistent phosphors allows sensitive analyte detection with a smartphone by a facile time-gated imaging strategy. Phosphors are first briefly excited with the phone’s camera flash, followed by switching off the flash, and subsequent imaging of phosphor luminescence with the camera. Using this approach, we demonstrate detection of human chorionic gonadotropin using a lateral flow assay and the smartphone platform with strontium aluminate nanoparticles as reporters, giving a detection limit of ≈45 pg/mL (1.2 pM) in buffer. Time-gated imaging on a smartphone can be readily adapted for sensitive and potentially quantitative testing using other point-of-care formats, and is workable with a variety of persistent luminescent materials.

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“…It was reported that oriented immobilization of antibodies increased the binding efficacy of a ligand by approximately 2-fold compared with that by random immobilization using amine-coupling reactions 17 .…”

Section: Construction Of a Magnetic Label By Oriented Immobilization mentioning

confidence: 99%

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

et al. 2011

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Early detection of human cardiac markers is regarded as a gold standard for diagnosis of acute myocardial infarction (AMI). Here we demonstrate the increase in the detection sensitivity of a lateral flow assay for a cardiac marker troponin I (cTnI) by oriented immobilization of antibodies on magnetic beads. For the construction of sensitive magnetic labels, orientated-immobilization of anti-troponin I antibodies on magnetic beads was attempted using protein G. Magnetic beads were first conjugated with protein G followed by immobilization of anti-cTnI antibodies. The factors affecting the detection sensitivity in the lateral flow assay system were investigated. The lateral flow assay of cTnI was carried out using the magnetic beads and a Giant magnetoresistive (GMR) sensor at the optimized conditions, and compared the results with using a conventional random immobilization method. Magnetic labels with oriented immobilization of antibodies enabled the detection of cTnI up to 0.01 ng/mL, showing significantly increased sensitivity compared with conventional immobilization method. This result clearly shows that the magnetic labels with oriented immobilization of antibodies can find wide applications in the lateral flow assays of target analytes.

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Improving immunobinding using oriented immobilization of an oxidized antibody

Cited by 45 publications

References 25 publications

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

A low-cost smartphone-based platform for highly sensitive point-of-care testing with persistent luminescent phosphors

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

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