The Effect of Length and Structure of Attached Polyethylene Glycol Chain on Hydrodynamic Radius, and Separation of PEGylated Human Serum Albumin by Chromatography

Akbarzadehlaleh, Parvin; Mirzaei, Mona; Mashahdi-Keshtiban, Mahdiyeh; Heidari, Hadi

doi:10.34172/apb.2021.082

Cited by 6 publications

(2 citation statements)

References 34 publications

(78 reference statements)

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“…To confirm such a trend, the hydrodynamic radius of the protein was measured by DLS (Figure ). The hydrodynamic radius of BSA monomer is about 3.3 nm in PBS and 4.1 nm in its dimeric form. − The mean radius of BSA in samples prepared at 0.25 mM was 4.5 nm, meaning that mostly dimers are present even if the width of the curves accounts for both monomers and larger oligomers. A slight increase for solutions with 0.5 mM BSA was obtained with an average value at 6.5 nm.…”

Section: Resultsmentioning

confidence: 99%

Kinetic Study of the Esterase-like Activity of Albumin following Condensation by Macromolecular Crowding

Lamy,

Bullier-Marchandin,

Pointel

et al. 2024

Biomacromolecules

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The ability of bovine serum albumin (BSA) to form condensates in crowded environments has been discovered only recently. Effects of this condensed state on the secondary structure of the protein have already been unraveled as some aging aspects, but the pseudoenzymatic behavior of condensed BSA has never been reported yet. This article investigates the kinetic profile of para-nitrophenol acetate hydrolysis by BSA in its condensed state with poly(ethylene) glycol (PEG) as the crowding agent. Furthermore, the initial BSA concentration was varied between 0.25 and 1 mM which allowed us to modify the size distribution, the volume fraction, and the partition coefficient (varying from 136 to 180). Hence, the amount of BSA originally added was a simple way to modulate the size and density of the condensates. Compared with dilute BSA, the initial velocity (v i ) with condensates was dramatically reduced. From the Michaelis−Menten fits, the extracted Michaelis constant K m and the maximum velocity V max decreased in control samples without condensates when the BSA concentration increased, which was attributed to BSA self-oligomerization. In samples containing condensates, the observed v i was interpreted as an effect of diluted BSA remaining in the supernatants and from the condensates. In supernatants, the crowding effect of PEG increased the k cat and catalytic efficiency. Last, V max was proportional to the volume fraction of the condensates, which could be controlled by varying its initial concentration. Hence, the major significance of this article is the control of the size and volume fraction of albumin condensates, along with their kinetic profile using liquid−liquid phase separation.

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Section: Resultsmentioning

confidence: 99%

Kinetic Study of the Esterase-like Activity of Albumin following Condensation by Macromolecular Crowding

Lamy,

Bullier-Marchandin,

Pointel

et al. 2024

Biomacromolecules

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show abstract

“…The reduction in hydrodynamic diameters could be attributed to the formation of a PEG shield, which effectively prevents interparticle aggregation. [ 34 ] Furthermore, zeta potential ( ζ ) measurements indicated a change in surface charge due to the polymer coatings, with values of −17.0 ± 0.4 and −17.3 ± 0.4 mV observed for Pd@MIL‐100(Fe)/PEG‐p(Asp) and Pd@MIL‐100(Fe)/PEG‐p(Asp‐Dopa), respectively, compared to −28.3 ± 0.7 mV for Pd@MIL‐100(Fe) (Table 1). The lower ζ ‐potential indicated the formation of a PEG shield.…”

Section: Resultsmentioning

confidence: 99%

Block Copolymer‐Stabilized Metal–Organic Framework Hybrids Loading Pd Nanoparticles Enable Tumor Remission Through Near‐Infrared Photothermal Therapy

Li,

Hsieh,

Hong

et al. 2023

Advanced NanoBiomed Research

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Metal–organic frameworks (MOFs), such as the magnetic resonance imaging‐fit MIL‐100 based on Fe, are gaining significant attention as versatile theranostics with high‐loading capability. Moreover, as MOFs can be engineered to target tumors, there is much interest in applying them for precise pin‐point treatment of cancer. Herein, Pd nanoparticles within MIL‐100(Fe) are generated to create MOFs with remarkable photothermal conversion properties for cancer therapy. The Pd‐loaded MIL‐100(Fe) (Pd@MIL‐100(Fe)) are stabilized with biocompatible block copolymers to generate MOFs with PEGylated surfaces. This is achieved by directly mixing poly(ethylene glycol)‐poly(L‐aspartic acid) (PEG‐p(Asp)) or dopamine‐modified PEG‐p(Asp) (PEG‐p(Asp‐Dopa)) block copolymers with the MOFs in aqueous conditions. The resulting block copolymer‐stabilized MOF hybrids are stable in physiological conditions. Particularly, the Pd@MIL‐100(Fe)/PEG‐p(Asp‐Dopa) hybrids show enhanced blood circulation and increased accumulation in B16F10 melanoma. Furthermore, when irradiated with 808 nm light, the Pd@MIL‐100(Fe)/PEG‐p(Asp‐Dopa) hybrids rapidly increase the temperature to 50 °C, enabling tumor remission. The surface‐stabilized Pd@MIL‐100(Fe)/polymer hybrids open viable opportunities for innovating MOF/polymer hybrid‐based approaches for drug delivery.

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Demonstration of Physicochemical and Functional Similarity of Biosimilar Pegfilgrastim-cbqv to Pegfilgrastim

Kuehne,

Davis,

Merewether

et al. 2024

Drugs R D

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Background Pegfilgrastim-cbqv/CHS-1701 (UDENYCA ® ) (hereafter referred to as pegfilgrastim-cbqv) was approved in 2018 by the US Food and Drug Administration as a biosimilar for pegfilgrastim (Neulasta ® ) (hereafter referred to as pegfilgrastim). Both pegfilgrastim-cbqv and pegfilgrastim are conjugates of recombinant human granulocyte colony stimulating factor (r-metHuG-CSF) with a 20 kDa polyethylene glycol (PEG) indicated to decrease the incidence of infection, as manifested by febrile neutropenia, in patients receiving myelosuppressive anticancer drugs. The demonstration of analytical similarity for PEG-protein conjugates presents unique challenges since both the protein and PEG attributes must be characterized. Objective The current study demonstrates the analytical similarity of pegfilgrastim-cbqv and the reference product, pegfilgrastim. In addition to the physicochemical and functional characterization of the protein, the study assessed attributes specific to PEGylation including PEG size and polydispersity, site of attachment, linker composition, and PEGylation process-related variants. Methods The structural, functional, and stability attributes of pegfilgrastim-cbqv and pegfilgrastim were compared using state-of-the-art analytical methods. For the protein, the primary structure, disulfide structure, and secondary and tertiary structures were assessed using traditional protein characterization techniques such as mass spectrometry (MS), circular dichroism (CD), intrinsic fluorescence, and differential scanning calorimetry (DSC), as well as more advanced techniques such as two-dimensional (2D) nuclear magnetic resonance (NMR) and hydrogen deuterium exchange (HDX). For the PEG moiety, the site of attachment, occupancy, linker composition, size and polydispersity were compared using mass spectrometry (both intact and after endoprotease digestion), multiangle light scattering detection (MALS), and Edman degradation. Purity assessments included the assessment of both protein variants and PEGylation variants using chromatographic and electrophoretic analytical separation techniques. The functional similarity between pegfilgrastim-cbqv and pegfilgrastim was compared using both a cell-based bioassay and surface plasmon resonance (SPR). The degradation rates and stability profiles were compared under accelerated and stressed conditions. Results Biosimilarity was demonstrated by a thorough assessment of physiochemical and functional attributes, as well as comparative stability, of pegfilgrastim-cbqv relative to pegfilgrastim. These studies demonstrated identical primary structure and disulfide structure, highly similar secondary and tertiary structure, as well as functional similarity. The impurity profile of pegfilgrastim-cbqv was comparable to that of pegfilgrastim with only minor differences in PEGylation variants and a slight offset in the PEG molar mass...

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The Effect of Length and Structure of Attached Polyethylene Glycol Chain on Hydrodynamic Radius, and Separation of PEGylated Human Serum Albumin by Chromatography

Cited by 6 publications

References 34 publications

Kinetic Study of the Esterase-like Activity of Albumin following Condensation by Macromolecular Crowding

Kinetic Study of the Esterase-like Activity of Albumin following Condensation by Macromolecular Crowding

Block Copolymer‐Stabilized Metal–Organic Framework Hybrids Loading Pd Nanoparticles Enable Tumor Remission Through Near‐Infrared Photothermal Therapy

Demonstration of Physicochemical and Functional Similarity of Biosimilar Pegfilgrastim-cbqv to Pegfilgrastim

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