Role of the Methoxy Group in Immune Responses to mPEG-Protein Conjugates

Sherman, Merry R.; Williams, L. David; Sobczyk, Monika; Michaels, Shawnya; Saifer, Mark G.P.

doi:10.1021/bc200551b

Cited by 138 publications

(124 citation statements)

References 47 publications

(124 reference statements)

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“…59 In contrast to these findings, another recent study found that the methoxy group of PEG leads to immune responses; however, if replaced with a hydroxyl group, the immune response was not pronounced. 60 Moreover, nanoparticle size has a role in ABC regardless of PEG density and chain length. Nanoparticle sizes .…”

Section: Effect Of Different Peg Mw and Ph On Size And Zeta Potentialmentioning

confidence: 99%

Development and characterization of chitosan-PEG-TAT nanoparticles for the intracellular delivery of siRNA

Malhotra¹,

Tomaro-Duchesneau²,

Saha³

et al. 2013

IJN

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Recently, cell-penetrating peptides have been proposed to translocate antibodies, proteins, and other molecules in targeted drug delivery. The proposed study presents the synthesis and characterization of a peptide-based chitosan nanoparticle for small interfering RNA (siRNA) delivery, in-vitro. Specifically, the synthesis included polyethylene glycol (PEG), a hydrophilic polymer, and trans-activated transcription (TAT) peptide, which were chemically conjugated on the chitosan polymer. The conjugation was achieved using N-Hydroxysuccinimide-PEGmaleimide (heterobifunctional PEG) as a cross-linker, with the bifunctional PEG facilitating the amidation reaction through its N-Hydroxysuccinimide group and reacting with the amines on chitosan. At the other end of PEG, the maleimide group was chemically conjugated with the cysteine-modified TAT peptide. The degree of substitution on chitosan with PEG and on PEG with TAT was confirmed using colorimetric assays. The resultant polymer was used to form nanoparticles complexing siRNA, which were then characterized for particle size, morphology, cellular uptake, and cytotoxicity. The nanoparticles were tested in-vitro on mouse neuroblastoma cells (Neuro2a). Particle size and surface charge were characterized and an optimal pH condition and PEG molecular weight were determined to form sterically stable nanoparticles. Results indicate 7.5% of the amines in chitosan polymer were conjugated to the PEG and complete conjugation of TAT peptide was observed on the synthesized PEGylated chitosan polymer. Compared with unmodified chitosan nanoparticles, the nanoparticles formed at pH 6 were monodispersed and of ,100 nm in size, exhibiting maximum cell transfection ability and very low cytotoxicity. Thus, this research may be of significance in translocating biotherapeutic molecules for intracellular delivery applications.

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Section: Effect Of Different Peg Mw and Ph On Size And Zeta Potentialmentioning

confidence: 99%

Development and characterization of chitosan-PEG-TAT nanoparticles for the intracellular delivery of siRNA

Malhotra¹,

Tomaro-Duchesneau²,

Saha³

et al. 2013

IJN

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“…Several studies have claimed that the PEG moiety of PEGylated therapeutic proteins itself might be immunogenic and could induce anti-PEG antibodies [27]. Further researches have indicated that the methoxy group within methoxy-PEG modified proteins played a critical role in the development of anti-PEG antibody with high affinity [27].…”

Section: Discussionmentioning

confidence: 99%

Effect of site-specific PEGylation on the fibrinolytic activity, immunogenicity, and pharmacokinetics of staphylokinase

Liu¹,

Wang²,

He³

et al. 2014

ABBS

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The bacterial plasminogen-activator staphylokinase (Sak) is a promising thrombolytic agent for treating acute myocardial infarction. To effectively reduce the immunogenicity of Sak while maintaining its fibrinolytic activity, site-specific PEGylation was performed in the present study. The chemoselective cysteine PEGylation site was selected within an immunodominant region (amino acid residues 71-87) using an in silico approach. The PEGylated Sak variants prepared in this study showed a purity of >97.0%. PEGylation at Position 80 resulted in a Sak variant Sak(E80C-PEG) which has similar fibrinolytic activity and thermostability compared with the native recombinant staphylokinase (r-Sak). The immunogenicity of Sak(E80C-PEG) in guinea pigs was greatly reduced compared with the native r-Sak. Furthermore, preliminary pharmacokinetic results suggested that the plasma clearance of Sak(E80C-PEG) from the blood stream of rabbit was significantly decreased compared with that of r-Sak, resulting in a 2.8-fold increase of initial half-life and a 3.8-fold increase of systemic availability. In summary, these results demonstrated that site-specific PEGylation yielded a novel Sak variant Sak(E80C-PEG) with remarkable advantages over the unmodified Sak.

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“…For this particular format, Casein in PBS from Thermo Fisher Scientific provided the lowest signal for the (14,15) previously reported on the detrimental effects of such surfactants on the sensitivity of detection for anti-PEG antibodies. Since our positive control antibody is PEG backbone unit specific, the interference was expected.…”

Section: Assay Buffermentioning

confidence: 99%

Development of a Generic Anti-PEG Antibody Assay Using BioScale’s Acoustic Membrane MicroParticle Technology

Dong

Mora

Brockus

et al. 2015

AAPS J

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Abstract. Immunogenicity testing for PEGylated biotherapeutics should include methods to detect both anti-protein and anti-PEG antibodies (anti-PEG). Although some methods have been published for the detection of anti-PEG antibodies, the information is incomplete and, in some cases, reagents used (such as Tween-20) are known to interfere with detection. This rapid communication describes the use of BioScale's Acoustic Membrane MicroParticle (AMMP®) technology using the ViBE® Workstation to measure anti-PEG antibodies in human serum samples. Briefly, a sample spiked with monoclonal human IgG anti-PEG antibody is diluted in buffer and incubated with paramagnetic beads coated with linear chain mPEG to capture anti-PEG antibodies. The complex is then captured on an acoustic membrane coated with Protein A. The change in mass on the membrane caused by the binding of the complex to the membrane results in a signal proportional to the mass of anti-PEG antibodies. The data indicate that an assay with a sensitivity of less than 1000 ng/mL for IgG is achievable. This level of sensitivity is better than current published reports on IgG anti-PEG antibody detection.

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Role of the Methoxy Group in Immune Responses to mPEG-Protein Conjugates

Cited by 138 publications

References 47 publications

Development and characterization of chitosan-PEG-TAT nanoparticles for the intracellular delivery of siRNA

Development and characterization of chitosan-PEG-TAT nanoparticles for the intracellular delivery of siRNA

Effect of site-specific PEGylation on the fibrinolytic activity, immunogenicity, and pharmacokinetics of staphylokinase

Development of a Generic Anti-PEG Antibody Assay Using BioScale’s Acoustic Membrane MicroParticle Technology

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