Surface plasmon resonance (SPR) based binding studies of refolded single chain antibody fragments

Singh, Pranveer

doi:10.1016/j.bbrep.2018.04.005

Cited by 5 publications

(3 citation statements)

References 46 publications

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“…[36] . Singh [37] showed that Fab with lower K D value evinced better binding (1.5 fold higher) with the studied protein as compared to scFv. In addition, the results reported by Maruta et al [38] suggested that the binding activity of antibody fragments, especially scFv (higher K D ), might be slightly reduced in comparison to that of whole antibodies.…”

Section: Discussionmentioning

confidence: 97%

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Karam

Raigani

Afshar

et al. 2021

ibj

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Section: Discussionmentioning

confidence: 97%

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Karam

Raigani

Afshar

et al. 2021

ibj

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“…Compared to the Fab format, scFv with a smaller molecular size has proper folding and assembly in the periplasm of E. coli . The Fab format also tends to form homo-dimers, which leads to decreased solubility and reduced expression level of Fab in E. coli [ 32 , 33 ]. RBD as target molecules in the spike (S) protein are abundantly expressed on virus surfaces.…”

Section: Discussionmentioning

confidence: 99%

Development of a Single-Chain Variable Fragment of CR3022 for a Plasmonic-Based Biosensor Targeting the SARS-CoV-2 Spike Protein

et al. 2022

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Two years after SARS-CoV-2 caused the first case of COVID-19, we are now in the “new normal” period, where people’s activity has bounced back, followed by the easing of travel policy restrictions. The lesson learned is that the wide availability of accurate and rapid testing procedures is crucial to overcome possible outbreaks in the future. Therefore, many laboratories worldwide have been racing to develop a new point-of-care diagnostic test. To aid continuous innovation, we developed a plasmonic-based biosensor designed explicitly for portable Surface Plasmon Resonance (SPR). In this study, we designed a single chain variable fragment (scFv) from the CR3022 antibody with a particular linker that inserted a cysteine residue at the second position. It caused the linker to have a strong affinity to the gold surface through thiol-coupling and possibly become a ready-to-use bioreceptor toward a portable SPR gold chip without purification steps. The theoretical affinity of this scFv on spike protein was −64.7 kcal/mol, computed using the Molecular Mechanics Generalized Born Surface Area (MM/GBSA) method from the 100 ns molecular dynamics trajectory. Furthermore, the scFv was produced in Escherichia coli BL21(DE3) as a soluble protein. The binding activity toward Spike Receptor Binding Domain (RBD) SARS-CoV-2 was confirmed with a spot-test, and the experimental binding free energy of −10.82 kcal/mol was determined using portable SPR spectroscopy. We hope this study will be useful in designing specific and low-cost bioreceptors, particularly early in an outbreak when the information on antibody capture is still limited.

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“…SPR is a label-free biosensor technique for studying interactions between all classes of biomolecules and biochemical mechanisms in real time 29 . This is done in a label free environment, while the ligand of interest is immobilized on the surface of the sensor chip and solutions with different concentrations of the analyte can flow over it.…”

Section: Surface Plasmon Resonancementioning

confidence: 99%

Binding kinetics of monoclonal antibodies and fluoroquinolone derivatives

Boonserm

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Fluoroquinolones (FQs) are a group of antibiotics which have been extensively used against both Gram-negative and Gram-positive bacteria. Some of FQs antibiotics commonly used in livestock and fishery include norfloxacin, enrofloxacin, ciprofloxacin, and ofloxacin. However, misuse of these antibiotics may cause drug residues in food products. Therefore, to prevent consumers from getting residual antibiotics in food and the drug resistance of pathogens in humans. As a result, surveillance detection program of these drug residues must be in practice to ensure safety of the consumers in many countries. In general, detections based on immunological method include enzyme-linked immunosorbent assay (ELISA) has been widely used as a screening tool in food safety applications. In this research, sensitivity of four monoclonal antibodies, mAbs against norfloxacin (mAb Nor132 and mAb Nor155), and mAbs against enrofloxacin (mAb Enro44 and Enro48) were quantified by an antigen-captured indirect competitive ELISA. The sensitivity in term of 50% inhibition concentration (IC??) of mAb Nor132, mAb Nor155, mAb Enro44, and mAb Enro48 were found to be 0.1388 ?g/ml, 0.0511 ?g/ml, 0.2369 ?g/ml, and 0.3320 ?g/ml respectively. Moreover, the specificity of mAbs were examined in term of % cross-reactivity. The mAb Nor132 and mAb Nor155 are highly specificity for norfloxacin and several drugs in FQs group in range of 21.79-89.34% but mAb Enro44 and mAb Enro48 are highly specificity only enrofloxacin. Then, the kinetic binding (KD) of four mAbs and drugs were studied by surface plasmon resonance (SPR). The result showed that mAb Nor155 bound to norfloxacin and ciprofloxacin better than mAb Nor132 did. As mAb Enro44 bound to enrofloxacin better than mAb Enro48. To investigate In silico binding between FQ drugs and mAbs by using antigen-antibody docking technique, Autodock vina program. The docking results expose significant interactions between the FQ dugs and four mAbs, especially with amino acid residues at the pocket site or nearby the area CDRs of mAbs, the region of mAb recognizes the antigen, with hydrogen bonds. The results indicated that sensitivity and affinity of mAb Nor155 were higher than those of mAb Nor132 as the mAb Enro44 compared to the mAb Enro48. These suggested that mAb Nor155 and mAb Enro44 were suitable for using in the development of norfloxacin and enrofloxacin detection in food products, respectively.

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Surface plasmon resonance (SPR) based binding studies of refolded single chain antibody fragments

Cited by 5 publications

References 46 publications

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Development of a Single-Chain Variable Fragment of CR3022 for a Plasmonic-Based Biosensor Targeting the SARS-CoV-2 Spike Protein

Binding kinetics of monoclonal antibodies and fluoroquinolone derivatives

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