Silica resins and peptide ligands to develop disposable affinity adsorbents for antibody purification

Naik, Amith D.; Islam, Tuhidul; Terasaka, Takaaki; Ohara, Yuki; Hashimoto, Yasuhiro; Menegatti, Stefano; Carbonell, Ruben G.

doi:10.1016/j.bej.2018.07.011

Cited by 6 publications

(4 citation statements)

References 47 publications

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“…The effluent was apportioned in 3-mL fractions, which were analyzed as described in Sections 4.12.2 and 4.12.3 to measure the titer of lentiviral genomes and transducing particles contained therein. The dynamic binding capacity at 10% of breakthrough (DBC 10% ) was calculated as described in prior work (Kish et al, 2017;Naik et al, 2019;Reese et al, 2020;Sripada et al, 2022;Xiao et al, 2022); the void volume of the system was measured via acetone pulse injection and utilized to adjust the value of DBC 10% . Sections 4.12.1 and 4.12.4 to measure LV yield and purity.…”

Section: Amination and Peptide Conjugation Of Poros™ 50 Oh Resinmentioning

confidence: 99%

Peptide ligands targeting the vesicular stomatitis virus G (VSV‐G) protein for the affinity purification of lentivirus particles

Barbieri,

Mollica,

Moore

et al. 2023

Biotech & Bioengineering

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The recent uptick in the approval of ex vivo cell therapies highlights the relevance of lentivirus (LV) as an enabling viral vector of modern medicine. As labile biologics, however, LVs pose critical challenges to industrial biomanufacturing. In particular, LV purification—currently reliant on filtration and anion‐exchange or size‐exclusion chromatography—suffers from long process times and low yield of transducing particles, which translate into high waiting time and cost to patients. Seeking to improve LV downstream processing, this study introduces peptides targeting the enveloped protein Vesicular stomatitis virus G (VSV‐G) to serve as affinity ligands for the chromatographic purification of LV particles. An ensemble of candidate ligands was initially discovered by implementing a dual‐fluorescence screening technology and a targeted in silico approach designed to identify sequences with high selectivity and tunable affinity. The selected peptides were conjugated on Poros resin and their LV binding‐and‐release performance was optimized by adjusting the flow rate, composition, and pH of the chromatographic buffers. Ligands GKEAAFAA and SRAFVGDADRD were selected for their high product yield (50%–60% of viral genomes; 40%–50% of HT1080 cell‐transducing particles) upon elution in PIPES buffer with 0.65 M NaCl at pH 7.4. The peptide‐based adsorbents also presented remarkable values of binding capacity (up to 3·109 TU per mL of resin, or 5·1011 vp per mL of resin, at the residence time of 1 min) and clearance of host cell proteins (up to a 220‐fold reduction of HEK293 HCPs). Additionally, GKEAAFAA demonstrated high resistance to caustic cleaning‐in‐place (0.5 M NaOH, 30 min) with no observable loss in product yield and quality.

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Section: Amination and Peptide Conjugation Of Poros™ 50 Oh Resinmentioning

confidence: 99%

Peptide ligands targeting the vesicular stomatitis virus G (VSV‐G) protein for the affinity purification of lentivirus particles

Barbieri,

Mollica,

Moore

et al. 2023

Biotech & Bioengineering

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“…The inorganic particles usually include porous silica particles and magnetic beads. Porous silica particles demonstrate a potential to be an alternative to traditional polymer supports on account of their easier regeneration, inexpensiveness, excellent flow properties, and easier surface modification [ 16 , 17 ]. Magnetic beads of different sizes are fabricated via entrapping magnetite within agarose, cellulose, polystyrene, or other polymeric materials, onto which ligands are fixed.…”

Section: Chromatographic Matrices and Corresponding Applications In R...mentioning

confidence: 99%

Matrices and Affinity Ligands for Antibody Purification and Corresponding Applications in Radiotherapy

Xue

Fan

2022

Biomolecules

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Antibodies have become an important class of biological products in cancer treatments such as radiotherapy. The growing therapeutic applications have driven a demand for high-purity antibodies. Affinity chromatography with a high affinity and specificity has always been utilized to separate antibodies from complex mixtures. Quality chromatographic components (matrices and affinity ligands) have either been found or generated to increase the purity and yield of antibodies. More importantly, some matrices (mainly particles) and affinity ligands (including design protocols) for antibody purification can act as radiosensitizers or carriers for therapeutic radionuclides (or for radiosensitizers) either directly or indirectly to improve the therapeutic efficiency of radiotherapy. This paper provides a brief overview on the matrices and ligands used in affinity chromatography that are involved in antibody purification and emphasizes their applications in radiotherapy to enrich potential approaches for improving the efficacy of radiotherapy.

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“…Several approaches have been designed and typically utilized to isolate, purify, and concentrate therapeutic biomolecules in solutions and water. − These approaches employ activated carbon, gel filtration, Triton X-100 phase separation, ultrafiltration, and affinity chromatography with adsorbents , functionalized with l -histidine, polyethyleneimine (pEI), poly-ε-lysine, poly(γ-methyl l -glutamate), or polymyxin B (PMB). − The abovementioned technologies also employed for the removal of endotoxins are unsatisfactory due to nonspecificity, low adsorption capacity, and poor recovery of products. Endotoxins have been extracted using ultrafiltration by taking advantage of the size-exclusion removal technique , or by nonselective binding with anion-exchange chromatography. , Affinity chromatography is applicable for a wide range of target molecules, including proteins and pDNA using specific interactions between endotoxin and a ligand bound to a stationary phase .…”

Section: Introductionmentioning

confidence: 99%

Multifunctional Biofilter to Effectively Remove Toxins

Razdan

Adler

Barua

et al. 2020

ACS Appl. Bio Mater.

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The goal of this work is to synthesize a multifunctional biofilter and assess its efficiency to remove endotoxins, freshwater harmful algal bloom (FHAB)-derived cyanotoxins, and toxic heavy metal ions from water using polymeric nanoparticle (PolyBall) embedded in a soy protein isolate (SPI)-derived cellulose acetate (CA) biofilter. The presence of even a trace number of endotoxins in biopharmaceutical solutions can create a pyrogenic response in the human body by triggering the immune system’s signaling cascade. The purification of products usually involves sequential ion-exchange chromatography, gel filtration, and/or ultrafiltration that are complex and expensive and are associated with high (>40%) product loss. The water treatment plants use a series of standard cleansing procedures to diminish cyanotoxins; however, finding an effective and affordable technique remains a challenge. We have developed a PolyBall scavenger of endotoxins, based on nonporous poly-ε-caprolactone (PCL) polymer nanoparticles (PCL NPs) that are capable of removing endotoxins from protein solutions at high efficiency (>99%) while maintaining >99% protein recovery. In this work, we put our efforts incorporating PCL NPs (PolyBalls) in SPI and CA biofilters to achieve improved efficiency in removing endotoxins, cyanotoxins, and toxic heavy metal ions such as lead ions from contaminated solutions. Six different recombinant protein solutions of molecular weights varying from 14 to 341 kDa and with isoelectric points (pI) varying from 4.5 to 10.7 were spiked with 5.74 × 105 EU/mL of endotoxins. The contaminated protein solutions were passed through biofilters that showed >99% of endotoxin removal from the solutions while maintaining >99% protein recovery. The biofilter exhibited excellent capability in cyanotoxin separation from surface water collected from three different lake water. This is the first demonstration of soy protein plant-based portable filters with embedded biocompatible polymeric NPs, which provide strong adsorptive interactions for toxin removal from protein solutions and surface water.

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Silica resins and peptide ligands to develop disposable affinity adsorbents for antibody purification

Cited by 6 publications

References 47 publications

Peptide ligands targeting the vesicular stomatitis virus G (VSV‐G) protein for the affinity purification of lentivirus particles

Peptide ligands targeting the vesicular stomatitis virus G (VSV‐G) protein for the affinity purification of lentivirus particles

Matrices and Affinity Ligands for Antibody Purification and Corresponding Applications in Radiotherapy

Multifunctional Biofilter to Effectively Remove Toxins

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