Methods of protein surface PEGylation under structure preservation for the emulsion-based formation of stable nanoparticles

Abstract

We evaluated different methods for a high surface PEGylation of lysozyme. The resulting lipophilic enzymes can be used for the formation of stable nanoparticles.

Pharmaceutically modified subtilisins withstand acidic conditions and effectively degrade gluten in vivo

“…Larger MW (>10 kDa) PEG molecules tend to fold and occupy a large surface area of the protein, interfering with substrate binding ability 33,34 . Our modification using 5 kDa mPEG yielded a high conjugation efficiency (~55%), which is comparable to studies with other protein substrates in which the PEG modification efficiency ranged between 50–60% 32,35 .…”

Physicochemical properties of free and immobilized tyrosinase from different species of yam (Dioscorea spp)

“…Modification of serine protease by pegylation, leading to increase molecular weight and higher catalytic efficiency of the enzyme was reported by da-Silva-Freitas et al [ 35 ]. Radi et al [ 36 ] has reported this type of behavior for lysozyme modification with epoxy-methoxy polyethylene glycol resulting in increased molecular masses of 30–34 kDa. Gooding et al [ 37 ] has also reported that there may be discrepancy in molecular weight as a result of modification of tyrosinase during purification as a result of association with other compounds.…”

Protein‐Based Nanoparticles for the Delivery of Enzymes with Antibacterial Activity

“…In order to obtain a suitable biopolymer material that can be applied for the double emulsion particle preparation, we had to change the solubility behavior of our chosen protein base material. The necessary surface modification of LYZ was performed, as previously described in our group, by the reaction of electrophilic‐activated 2,3,5,6‐tetrafluorophenyl 4‐((2‐methoxypolyethylene glycol)amino)‐4 oxobutanoate (TFP‐mPEG) with free amines on the surface of lysozyme …”

“…However, all of the mentioned particle preparation methods usually contain steps that require the denaturation of the proteins or permanent chemical cross‐linking to obtain stable nanoparticles. Our group described previously a single emulsion method of protein‐based nanoparticles for the encapsulation of hydrophobic drugs like doxorubicin, resulting in stable nanoparticles without the addition of a cross‐linking agent . For this, a single emulsion method was applied, where the hydrophobic payload is dissolved in organic solvents together with the lipophilic particle material before being emulsified with water.…”