PEGylation Analysis and Method Verification

Name: PEGylation Analysis and Method Verification
Brand: BOC Sciences

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BOC Sciences provides comprehensive PEGylation analysis services in terms of qualitative assessment, quantitative assessment and in vivo & in vitro characterization.

After PEGylation, conjugates will show changes in physiochemical properties, such as surface charge, hydrodynamic diameter, hydrophilicity and protein binding capability. By monitoring changes in these physiochemical properties, PEGylation can be confirmed.

Qualitative Assessments

Method	Analysis Content
Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE)	Purity of PEGylated proteins and the extent of PEGylation
Dynamic Light Scattering (DLS)	Hydrodynamic diameter, zeta potential and size distribution of PEGylated species
High Performance Size Exclusion Chromatography (HPSEC)	The size of PEGylated proteins
Circular Dichroism Measurement (CD)	Secondary and tertiary structural conformation of native and PEGylated proteins.
Fourier transform-infrared spectroscopy (FT-IR)	Qualitative assessment of PEGylation relies on the presence of CH2 and C-O-C absorption bands.
Zeta Potential Measurement	The surface charge of PEGylated species
13C-Nuclear Magnetic Resonance (13C-NMR)	Qualitative assessment of PEGylation relies on the presence or absence of specific chemical moieties
1H-Nuclear Magnetic Resonance (1H-NMR)	Qualitative assessment of the structural conformations of the lipid PEGylated nanocarriers
Transmission Electron Microscopy (TEM)	The morphology of PEGylated nanocarriers
Atomic Force Microscopy (AFM)	Identification the density and spatial distribution of PEG polymers on the surface of nanocarriers
Scanning Tunneling Microscopy (STM)	Identification the density and spatial distribution of PEG polymers on the surface of nanocarriers
Gel Permeation Chromatography (GPC)	Evaluation of the degree of PEG substitution
Many More …

Quantitative Assessments

Method	Analysis Content
High Performance Liquid Chromatography (HPLC)	The amount of PEG associated with colloidal carriers
Enzyme-linked Immunosorbent Assay (ELISA)	The concentration of PEGylated proteins in serum samples
Colorimetric Assays	The concentration and localization of PEG chains
Gas Chromatography (GC)	The amount of attached PEG polymer
Mass Spectrometry (MS)	The molecular mass of PEGylated species
Liquid Chromatography Mass Spectrometry (LC-MS)	The molecular mass of PEGylated species
Matrix-Assisted Laser Desorption Ionization Mass Spectrometry (MALDI–MS)	The average molecular weight of PEGylated species and degree of PEGylation
Electrospray Ionization Mass Spectrometry (ESI-MS)	The average molecular weight of PEGylated species and degree of PEGylation
X-Ray Photoelectron Spectroscopy (XPS)	Changes of elemental composition on the conjugate surface before and after PEGylation
Raman Spectroscopy	Determination of PEG content and conformation
Thermogravimetric analysis (TGA)	The content of PEG
Inductively Coupled Plasma Mass Spectrometry (ICP-MS)	The quantitative measurement of metal-containing PEGylated pharmaceuticals or nanoparticles
Many More …

In addition to using the analytical measurement mentioned above alone, we are also able to utilize the combined techniques, such as combination of ELSD and LC-MS, NIR spectroscopy coupled with principal component analysis (PCA) for characterization of hydrophilic/hydrophobic balance of PEGylated nanoparticles.

In Vitro and In Vivo Characterizations

In vitro methods include measurement of protein adsorption, measurement of complement consumption, macrophage uptake studies, and the analysis of release rate or content stability.
The most general in vivo assessments include pharmacokinetic profiling (Radiolabeling, Fluorescence labeling, Bioactivity Assay), tissue distribution studies and vascular permeability studies.

Why Us?

Advantage of BOC Sciences

How to Order?

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Reference

Santos, J. H. P. M.; Torres-Obreque, K. M.; et al. Protein PEGylation for the design of biobetters: from reaction to purification processes. Brazilian Journal of Pharmaceutical Sciences 2018, 54 (SPE).

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