Ion-exchange resins facilitate like-charged protein refolding: Effects of porous solid phase properties

Yu, Linling; Dong, Xiaoyan; Sun, Yan

doi:10.1016/j.chroma.2011.12.078

Cited by 26 publications

(3 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Other possibility is the modification of the support, e.g., adding anion groups [60], or even mixed cation/anion groups [61]. Some authors suggest that enzyme refolding of immobilized enzymes is easier if the support has the same ion nature of the enzyme, this may help in improving enzyme activity recovery during activity determination under mild conditions [62,63,64,65,66].…”

Section: Discussionmentioning

confidence: 99%

Reversible Immobilization of Lipases on Heterofunctional Octyl-Amino Agarose Beads Prevents Enzyme Desorption

et al. 2016

View full text Add to dashboard Cite

Abstract:Two different heterofunctional octyl-amino supports have been prepared using ethylenediamine and hexylendiamine (OCEDA and OCHDA) and utilized to immobilize five lipases (lipases A (CALA) and B (CALB) from Candida antarctica, lipases from Thermomyces lanuginosus (TLL), from Rhizomucor miehei (RML) and from Candida rugosa (CRL) and the phospholipase Lecitase Ultra (LU). Using pH 5 and 50 mM sodium acetate, the immobilizations proceeded via interfacial activation on the octyl layer, after some ionic bridges were established. These supports did not release enzyme when incubated at Triton X-100 concentrations that released all enzyme molecules from the octyl support. The octyl support produced significant enzyme hyperactivation, except for CALB. However, the activities of the immobilized enzymes were usually slightly higher using the new supports than the octyl ones. Thermal and solvent stabilities of LU and TLL were significantly improved compared to the OC counterparts, while in the other enzymes the stability decreased in most cases (depending on the pH value). As a general rule, OCEDA had lower negative effects on the stability of the immobilized enzymes than OCHDA and while in solvent inactivation the enzyme molecules remained attached to the support using the new supports and were released using monofunctional octyl supports, in thermal inactivations this only occurred in certain cases.

show abstract

Section: Discussionmentioning

confidence: 99%

Reversible Immobilization of Lipases on Heterofunctional Octyl-Amino Agarose Beads Prevents Enzyme Desorption

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Secondly, 2 g of pGMA‐OH (wet) was added to a mixture of 4 mL DMSO, 2 mL ECH, and 4 mL NaOH (1 M) for 2.5 h at 25°C and 170 rpm 28 . The epoxied microspheres were washed with distilled water to remove any other organic residue, and the product was denoted as pGMA‐ECH.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a heptapeptide‐modified microsphere for oriented antibody immobilization

Bai

Zhang

Zhu

et al. 2022

Journal of Peptide Science

Self Cite

View full text Add to dashboard Cite

Oriented immobilization of antibodies is important for the effective recognition of target antigens. In this paper, a heptapeptide ligand, HWRGWVC (HC7), was modified onto non-porous monosized poly(glyceryl methacrylate) (pGMA) microspheres (named pGMA-HC7) to explore the antibody immobilization behaviors. Characterization of the microspheres by particle size analyzer, scanning electron microscopy, Fourier transform infrared spectroscopy, and reversed-phase chromatography proved the success of each fabrication step. The capacity and activity of antibody immobilization through HC7 were studied using immunoglobulin G (IgG) as a model antibody and horseradish peroxidase (HRP) as a model antigen. Additionally, IgG immobilizations on pGMA microspheres by nonspecific adsorption and covalent coupling through carbodiimide chemistry were conducted for comparison. pGMA-HC7 exhibited an IgG adsorption capacity of 3-4 mg/g in 10 min by the specific binding of HC7 without nonspecific interactions. Notably, the ligand HC7 showed a by two orders of magnitude stronger affinity for IgG than its original hexapeptide ligand HWRGWV. Moreover, the capacity and activity of the immobilized anti-HRP antibody on pGMA-HC7 were 1.6-fold and 3-fold higher than those of the covalent coupling, respectively. The results proved the superior role of HWRGWVC in the affinity binding of antibody and the potential of pGMA-HC7-25 in immunoassay and immunodiagnostic applications.

show abstract

“…Investigating the influence of various surfaces on protein–protein interactions would be helpful but is usually overlooked. Electrostatic forces in solutions of like-charged colloidal particles were investigated, and electrostatic attraction was usually more focused. − For electrostatic repulsion, a successful regulation on protein–protein interactions using like-charged surfaces in vitro was experimentally found in our previous work, which facilitated the on-pathway folding of proteins expressed as inclusion bodies. − The working mechanism supposed was that the charged surfaces-oriented alignment of like-charged protein molecules inhibited protein–protein interactions. However, experimental verification of such a mechanism, especially the electrostatic repulsion-oriented alignment, was a challenge.…”

Section: Introductionmentioning

confidence: 99%

Charged Surface Regulates the Molecular Interactions of Electrostatically Repulsive Peptides by Inducing Oriented Alignment

Lin

Sun

2018

Langmuir

Self Cite

View full text Add to dashboard Cite

Regulation of molecular orientation of charged dipeptides and involved interactions by electrostatic repulsion from like-charged surfaces were studied using all-atom molecular dynamics simulations. It was found that a charged surface can induce oriented alignment of like-charged peptides, and the oriented alignment leads to enhanced electrostatic repulsion between the peptide molecules. The findings are consistent with previous experimental results about the inhibition of charged protein aggregation using like-charged ion-exchange resin. Furthermore, the simulations provided molecular insights into this process, and demonstrated the distinct regulation effect of like-charged surfaces on the molecular interactions between peptides that possess an electric dipole structure. Both the charged surface and the electric dipole structure of peptides were confirmed to be crucial for the regulation. The research is expected to facilitate the rational design of surfaces or devices to regulate the behavior of amphoteric molecules such as proteins for both in vivo and in vitro applications, which would contribute to the regulation of protein-protein interactions and its application in life science and biotechnology.

show abstract

Ion-exchange resins facilitate like-charged protein refolding: Effects of porous solid phase properties

Cited by 26 publications

References 39 publications

Reversible Immobilization of Lipases on Heterofunctional Octyl-Amino Agarose Beads Prevents Enzyme Desorption

Reversible Immobilization of Lipases on Heterofunctional Octyl-Amino Agarose Beads Prevents Enzyme Desorption

Characterization of a heptapeptide‐modified microsphere for oriented antibody immobilization

Charged Surface Regulates the Molecular Interactions of Electrostatically Repulsive Peptides by Inducing Oriented Alignment

Contact Info

Product

Resources

About