Antibody drug separation using thermoresponsive anionic polymer brush modified beads with optimised electrostatic and hydrophobic interactions

Nagase, Kenichi; Ishii, Saki; Ikeda, Kazuhiko; Yamada, Sota; Ichikawa, Daiju; Akimoto, Aya Mizutani; Hattori, Yutaka; Kanazawa, Hideko

doi:10.1038/s41598-020-68707-7

Cited by 37 publications

(15 citation statements)

References 53 publications

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“…Bead suspensions (0.5 mg/mL) were prepared in a 5 mmol/L potassium chloride solution for zeta potential measurements 59 . The morphology of the silica beads at each reaction step was observed using SEM (TM4000Plus-II, Hitachi High-tech, Tokyo, Japan) 41 .…”

Section: Methodsmentioning

confidence: 99%

“…Owing to its thermoresponsive properties, PNIPAAm has been used in various applications, such as drug and gene delivery systems 11 – 16 , biosensor and bioimaging systems 17 – 22 , nano-actuators 23 – 27 , cell separation systems 28 – 34 , and cell culture substrates 35 – 40 . In a previous study, a PNIPAAm-modified stationary phase was used in a chromatography system because of its hydrophobicity, which changes with temperature 41 – 45 . The hydrophobic interaction between the stationary phase and analytes can be modulated by changing the column temperature.…”

Section: Introductionmentioning

confidence: 99%

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Temperature-responsive mixed-mode column for the modulation of multiple interactions

Nagase

Matsumoto

Kanazawa

2022

Sci Rep

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In this study, mixed-mode chromatography columns have been investigated using multiple analyte interactions. A mixed-mode chromatography column was developed using poly(N-isopropylacrylamide) (PNIPAAm) brush-modified silica beads and poly(3-acrylamidopropyl trimethylammonium chloride) (PAPTAC) brush-modified silica beads. PNIPAAm brush-modified silica beads and PAPTAC brush-modified silica beads were prepared by atom transfer radical polymerization. The beads were then packed into a stainless-steel column in arbitrary compositions. The elution studies evaluated the column performance on hydrophobic, electrostatic, and therapeutic drug samples using steroids, adenosine nucleotide, and antiepileptic drugs as analytes, respectively. Steroids exhibited an increased retention time when the column temperature was increased. The retention of adenosine nucleotides increased with the increasing composition of the PAPTAC-modified beads in the column. The antiepileptic drugs were separated using the prepared mixed-mode columns. An effective separation of antiepileptic drugs was observed on a 10:1 PNIPAAm:PAPTAC column because the balance between the hydrophobic and electrostatic interactions with antiepileptic drugs was optimized for the bead composition. Oligonucleotides were also separated using mixed-mode columns through multiple hydrophobic and electrostatic interactions. These results demonstrate that the developed mixed-mode column can modulate multiple hydrophobic and electrostatic interactions by changing the column temperature and composition of the packed PNIPAAm and PAPTAC beads.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Temperature-responsive mixed-mode column for the modulation of multiple interactions

Nagase

Matsumoto

Kanazawa

2022

Sci Rep

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“…In contrast, the polymer transitioning to a hydrophilic state will promote protein desorption [ 18 , 24 ]. Both PVCL and PNIPAM have been investigated for application as HIC ligands for protein purifications [ 4 , 5 , 6 , 7 , 8 , 10 , 24 , 25 , 26 , 27 ]. However, the advantages of using PVCL are its biocompatibility and low toxicity.…”

Section: Introductionmentioning

confidence: 99%

Electrospun Hydrophobic Interaction Chromatography (HIC) Membranes for Protein Purification

2022

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Responsive membranes for hydrophobic interaction chromatography have been fabricated by functionalizing poly(N-vinylcaprolactam) (PVCL) ligands on the substrate of electrospun regenerated cellulose nanofibers. Both static and dynamic binding capacities and product recovery were investigated using bovine serum albumin (BSA) and Immunoglobulin G (IgG) as model proteins. The effects of ligand chain length and chain density on static binding capacity were also studied. A static binding capacity of ~25 mg/mL of membrane volume (MV) can be achieved in optimal ligand grafting conditions. For dynamic binding studies, protein binding capacity increased with protein concentration from 0.1 to 1.0 g/L. Dynamic binding capacity increased from ~8 mg/mL MV at 0.1 g/L BSA to over 30 mg/mL at 1.0 g/L BSA. However, BSA recovery decreased as protein concentration increased from ~98% at 0.1 g/L BSA to 51% at 1 g/L BSA loading concentration. There is a clear trade-off between binding capacity and recovery rate. The electrospun substrate with thicker fibers and more open pore structures is superior to thinner fibrous membrane substrates.

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“…Thermoresponsive polymers (TRP)s exhibit an abrupt volume phase transition by increasing temperature above their hydrophilic hydrophobic transition or lower critical solution temperature (LCST) . TRPs with their LCST close to biological temperature, such as poly( N -isopropylacrylamide) (pNIPAAm) and poly(vinyl methy lether) (PVME), are attractive candidates for a wide range of applications, including drug delivery, cell culture supports, protein chromatography, sensing devices, and regenerative medicine. − …”

Section: Introductionmentioning

confidence: 99%

Thermoresponsive Poly(vinyl methyl ether) (PVME) Retained by 3-Aminopropyltriethoxysilane (APTES) Network

Malekzadeh

Newby

2020

ACS Biomater. Sci. Eng.

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Thermoresponsive polymers (TRP)s have been widely used for various applications from controlling membrane fouling in separation to cell/cell sheet harvesting in regenerative medicine. While poly(N-isopropylacrylamide) (pNIPAAm) is the most commonly used TRP, less expensive and easily processed poly(vinyl methyl ether) (PVME) also shows a hydrophilic to hydrophobic transition at 32–35 °C, near physiological conditions. In this study, we investigated the processing conditions for retaining a stable layer of PVME thin film on silica surfaces via entrapment in a 3-aminopropyltriethoxysilane (APTES) network. In addition, the thermoresponsive behaviors (TRB) of the retained PVME films were evaluated. Blend thin films of PVME/APTES with 90:10 and 50:50 mass ratios were spin-coated from their solutions in ethanol under ambient conditions and then annealed in a vacuum oven at 40, 60, 80, or 120 °C for 1, 2, or 3 days. The annealed films were then thoroughly rinsed with room temperature water and then soaked in water for 3 days. Our results showed that annealing at a temperature of ≥40 °C was necessary for retaining a PVME film on the surface. The higher annealing temperature led to greater film retention, probably due to the formation of a tighter APTES network. Regardless of processing conditions, all retained PVME films showed TRB, determined by water contact angles below and above the transition temperature of PVME. Additionally, particle attachment and protein adsorption on retained PVME films showed lower attachment or adsorption at room temperature as compared to that at 37 °C, and a greater difference was observed for the 90:10 blend where more PVME was consisted. Furthermore, human mesenchymal stem cells attached and proliferated on the retained PVME surfaces at 37 °C and rapidly detached at room temperature. These results illustrated the potential applications of PVME surfaces as thermoresponsive supports for low-fouling applications and noninvasive cell harvesting.

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Antibody drug separation using thermoresponsive anionic polymer brush modified beads with optimised electrostatic and hydrophobic interactions

Cited by 37 publications

References 53 publications

Temperature-responsive mixed-mode column for the modulation of multiple interactions

Temperature-responsive mixed-mode column for the modulation of multiple interactions

Electrospun Hydrophobic Interaction Chromatography (HIC) Membranes for Protein Purification

Thermoresponsive Poly(vinyl methyl ether) (PVME) Retained by 3-Aminopropyltriethoxysilane (APTES) Network

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