Isolation of Positional Isomers of Mono-poly(ethylene glycol)ylated Interferon/α-2a and the Determination of Their Biochemical and Biological Characteristics

Monkarsh, Seth P.; Spence, Cheryl; Porter, Jill E.; Palleroni, Alicia V.; Nalin, Carlo M.; Rosen, Perry; Bailon, Pascal

doi:10.1021/bk-1997-0680.ch015

Cited by 29 publications

(9 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Initially, the protein was PEGylated via a urea linkage with an mPEG 5-kDa reagent. [99][100][101] A cation exchange method was developed to separate all 11 mono-PEGylated isoforms. Peptide mapping was used to verify that all 11 lysines on the native protein were PEGylated but not the N-terminus.…”

Section: Pegasys® (Peg Inf-a2a)mentioning

confidence: 99%

PEGylation of Biopharmaceuticals: A Review of Chemistry and Nonclinical Safety Information of Approved Drugs

Turecek¹,

Bossard

Schoetens

et al. 2016

Journal of Pharmaceutical Sciences

612

455

View full text Add to dashboard Cite

Modification of biopharmaceutical molecules by covalent conjugation of polyethylene glycol (PEG) molecules is known to enhance pharmacologic and pharmaceutical properties of proteins and other large molecules and has been used successfully in 12 approved drugs. Both linear and branched-chain PEG reagents with molecular sizes of up to 40 kDa have been used with a variety of different PEG derivatives with different linker chemistries. This review describes the properties of PEG itself, the history and evolution of PEGylation chemistry, and provides examples of PEGylated drugs with an established medical history. A trend toward the use of complex PEG architectures and larger PEG polymers, but with very pure and well-characterized PEG reagents is described. Nonclinical toxicology findings related to PEG in approved PEGylated biopharmaceuticals are summarized. The effect attributed to the PEG part of the molecules as observed in 5 of the 12 marketed products was cellular vacuolation seen microscopically mainly in phagocytic cells which is likely related to their biological function to absorb and remove particles and macromolecules from blood and tissues. Experience with marketed PEGylated products indicates that adverse effects in toxicology studies are usually related to the active part of the drug but not to the PEG moiety.

show abstract

Section: Pegasys® (Peg Inf-a2a)mentioning

confidence: 99%

PEGylation of Biopharmaceuticals: A Review of Chemistry and Nonclinical Safety Information of Approved Drugs

Turecek¹,

Bossard

Schoetens

et al. 2016

Journal of Pharmaceutical Sciences

612

455

View full text Add to dashboard Cite

show abstract

“…The identification of PEGylation sites in proteins is essentially needed to better characterize PEGylated proteins for the purpose of maximally retaining their biological activities after the PEG conjugation. The most widely known method in identifying PEGylation sites has been the Edman degradation, but this method cannot be applied for large peptides and proteins because of difficulty in purifying individual fragments produced after the enzymatic cleavage of the conjugates 9–11. Peptide mapping, a method based on enzymatic digestion followed by a reversed‐phase high‐pressure liquid chromatography (RP‐HPLC), has been widely used for identifying the PEGylation site by comparing digested chromatogram patterns between PEGylated and native proteins 12–15.…”

Section: Introductionmentioning

confidence: 99%

A Novel Method for Identifying PEGylation Sites of Protein Using Biotinylated PEG Derivatives

Lee

Park

2003

Journal of Pharmaceutical Sciences

View full text Add to dashboard Cite

“…Monkarsh et al (1997) used this technique to PEGylate α-interferon before separating the positional isomers. They bound the native protein to an ion exchanger and then passed activated PEG through the column.…”

Section: Reaction Engineeringmentioning

confidence: 99%

“…Monkarsh et al (1997) were able to separate 11 positional isomers of monoPEGylated α-interferon using cation exchange chromatography but only in analytical scale. Differences between isomers in isoelectric point (pI), surface charge distribution and relative hydrophobicity are likely to be subtle and it is doubtful that these can be exploited at the preparative scale.…”

Section: General Considerationsmentioning

confidence: 99%

PEG-proteins: Reaction engineering and separation issues

Fee

Alstine²

2006

Chemical Engineering Science

217

171

View full text Add to dashboard Cite

Poly(ethylene glycol)-conjugated (or PEGylated) proteins are an increasingly important class of therapeutic proteins that offer improved in vivo circulation half lives over their corresponding native forms. Their production involves covalent attachment of one or more poly(ethylene glycol) molecules to a native protein, followed by purification. Because of the extremely high costs involved in producing native therapeutic proteins it is important that subsequent PEGylation processes are as efficient as possible. In this paper, reaction engineering and purification issues for PEGylated proteins are reviewed. Paramount considerations for PEGylation reactions are specificity with respect to the conjugation site and overall yield. Batch PEGylation reaction methods are discussed, along with innovative methods using packed bed or "on-column" approaches to improve specificity and yield. Purification methods are currently dominated by ion exchange and size exclusion chromatography. Other methods in common use for protein separations, including hydrophobic interaction chromatography, affinity chromatography and membrane separations, are rarely used in PEGylated protein purification schemes. A better understanding of the effects of PEGylation on the physicochemical properties of proteins (isoelectric point, surface charge density and distribution, molecular size and relative hydrophobicity) and interactions between PEGylated proteins and surfaces is needed for the future development of optimal purification processes and media.

show abstract

Isolation of Positional Isomers of Mono-poly(ethylene glycol)ylated Interferon/α-2a and the Determination of Their Biochemical and Biological Characteristics

Cited by 29 publications

References 0 publications

PEGylation of Biopharmaceuticals: A Review of Chemistry and Nonclinical Safety Information of Approved Drugs

PEGylation of Biopharmaceuticals: A Review of Chemistry and Nonclinical Safety Information of Approved Drugs

A Novel Method for Identifying PEGylation Sites of Protein Using Biotinylated PEG Derivatives

PEG-proteins: Reaction engineering and separation issues

Contact Info

Product

Resources

About