Batch adsorption/desorption for purification of scFv antibodies using nanozeolite microspheres

Mesgari-Shadi, Ali; Sarrafzadeh, Mohammad‐Hossein; Divband, Baharak; Barar, Jaleh; Omidi, Yadollah

doi:10.1016/j.micromeso.2018.01.028

Cited by 13 publications

(7 citation statements)

References 49 publications

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“…A comparison of the IMAC resin designed in this study with the resins used in other studies for scFv antibody purification showed the high potential of the proposed resin. 22 The purity obtained with this method was over 90%. Sushma et al designed the IMAC resin with a copper ion stabilization on the commercial methacrylate substrate for purification of the scFv antibodies and obtained 90% purification after two-stage purification by IMAC resin and a commercial anion exchange column.…”

Section: Resultsmentioning

confidence: 95%

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Purification of a Novel Anti-VEGFR2 Single Chain Antibody Fragmentand Evaluation of Binding Affinity by Surface Plasmon Resonance

et al. 2019

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Purpose: The single-chain variable fragment (scFv) domain of antibodies is now considered asone of the therapeutic tools that can be produced by phage display technology (PDT). Antibodypurification is one of the most important steps in antibodies production. The aim of study waspurification and characterization of anti-VEGFR2 scFv antibody fragments.Methods: After the coating of vascular endothelial growth factor receptor 2 (VEGFR2) peptidein ELISA microplates, the phage display library of Tomlinson was used for antibody isolation.The targeted scFv was purified by chromatography using a zeolite-based column. The purity andfunctional assessment of purified scFv were evaluated by sodium dodecyl sulfate polyacrylamidegel electrophoresis (SDS-PAGE) and western blotting techniques, respectively. Affinity bindingwas evaluated by surface plasmon resonance (SPR).Results: The desired scFv was selected after four stages of biopanning. SDS-PAGE analysisshowed a 28 kDa scFv with high purity (>90%). The western bloting analysis confirmed thebinding of produced scFv antibody to the desired peptide. The affinity binding of scFv antibodyanalyzed by SPR was about 60 μM.Conclusion: In this study, the novel scFv antibody against VEGFR2 peptide was purified bychromatography column containing zeolite. Based on our findings the produced antibody maybe applied for diagnosis or targeting of VEGFR2 in antibody-based therapy strategies.

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Section: Resultsmentioning

confidence: 95%

“…After selection of scFv by biopanning with phage display technology (PDT) and its expression in host, the purification of scFv antibodies was performed using zeolite (ZSM-5-Ni +2 ) using the BOECO Rotator Multi Bio RS-24. 22…”

Section: Methodsmentioning

confidence: 99%

Purification of a Novel Anti-VEGFR2 Single Chain Antibody Fragmentand Evaluation of Binding Affinity by Surface Plasmon Resonance

et al. 2019

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“…In this in vitro study, the 13X zeolite was prepared based on the study carried out by Mesgari-Shadi et al [20]. It was dispersed in 2 ml distilled water using an ultrasound device for 2 h. Then, gadolinium chloride solution with a ratio of 2.6 wt% zeolite was added to the suspension, stirred at room temperature for 24 h with a magnetic stirrer and centrifuged.…”

Section: Preparation Of Gd 3+ /13x/fa/dox Nanocompositementioning

confidence: 99%

Magnetic Resonance Imaging Property of Doxorubicin-Loaded Gadolinium/13X Zeolite/Folic Acid Nanocomposite

Ghaderi

2020

J Biomed Phys Eng

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Background: Magnetic resonance imaging (MRI) using nanostructures has been a proper method for tumor targeting purposes. Different MRI nanomaterials, targeting agents and anticancer drugs have been used for targeting of tumors. Objectives: This study aims to consider the MRI property of doxorubicin (DOX)-loaded gadolinium/13X zeolite/folic acid (Gd3+/13X/FA) nanocomposite. Material and Methods: In this in vitro study, Gd3+/13X/FA/DOX nanocomposite was prepared and the X-ray diffraction, scanning electron microscopy and MTT assay were conducted to evaluate the physicochemical properties of the nanocomposite. MRI was performed at 25°C using a 1.5 T clinical system to determine the T1 relaxation times and subsequently, the T1 relaxivity. Results: The size of the nanocomposite was in the range of 80-200 nm. The nanocomposite without DOX loading (Gd3+/13X/FA) showed compatibility for A549 cells for all concentrations while DOX-loaded nanocomposite was toxic for 62% of the cells at the concentration of 0.4 mg/ml. The T1 relaxivity of Gd3+/13X/FA/DOX nanocomposite was 4.0401 mM-1s-1. Conclusion: Gd3+/13X/FA/DOX nanocomposite shows a T1 relaxivity similar to the conventional gadolinium chelates, and a successful DOX loading.

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“…Two new types of nanozeolite microspheres have been described . Zeolites are aluminosilicates having well‐defined inner structure.…”

Section: Specialtiesmentioning

confidence: 99%

Recent advances in mixed‐mode chromatographic stationary phases

Sýkora

Řezanka

Záruba

et al. 2018

J of Separation Science

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Mixed‐mode phases have become very popular in the last decade, and the number of new mixed/multi‐mode sorbents is growing fast. Unlike single‐mode stationary phases, perfectly suited for the separation of the analytes possessing similar physicochemical properties, for instance reversed‐phase chromatography for hydrophobic solutes, mixed‐mode sorbents providing multimodal interactions can render better separation selectivity for complex mixtures of solutes differing significantly in their physicochemical characteristics. The most frequent modern mixed‐mode stationary phases are di/tri‐mode sorbents embracing the following interactions, hydrophobic, electrostatic (coulombic), and hydrophilic. According to their structures, it is possible to distinguish silica‐based, polymer‐based, hybrid, and monolithic mixed‐mode stationary phases. Herewith, newly synthesized mixed‐mode sorbents developed within the last two and half years are categorized, discussed, and summarized. The main attention is devoted to the description of the synthetic routes and characterization methods applied for the new stationary phases.

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Batch adsorption/desorption for purification of scFv antibodies using nanozeolite microspheres

Cited by 13 publications

References 49 publications

Purification of a Novel Anti-VEGFR2 Single Chain Antibody Fragmentand Evaluation of Binding Affinity by Surface Plasmon Resonance

Purification of a Novel Anti-VEGFR2 Single Chain Antibody Fragmentand Evaluation of Binding Affinity by Surface Plasmon Resonance

Magnetic Resonance Imaging Property of Doxorubicin-Loaded Gadolinium/13X Zeolite/Folic Acid Nanocomposite

Recent advances in mixed‐mode chromatographic stationary phases

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