PEGylation of human serum albumin

Reaction of PEG-phenyl-isothiocyanate with protein

Fantao Meng, Belur N. Manjula, Paul K. Smith, Seetharama A. Acharya

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Successful and cost-effective PEGylation protocols require pure functionalized PEG reagents, which can be synthesized by simple and efficient procedures, exhibit high stability against hydrolysis, and maintain a level of reactivity with protein functional groups under mild reaction conditions. PEG-phenyl-isothiocyanate (PIT-PEG) is a new functionalized PEG having these characteristics, and has been synthesized by condensation of the bifunctional reagent 4-isothiocyanato phenyl isocyanate with monomethoxy PEG (mPEG). The data of 1H NMR and colormetric analysis of the new PEG reagent establish that the mPEG has been quantitatively functionalized. The t1/4 values for the hydrolysis of PIT-PEG5K in 100 mM phosphate solution at pH 6.5 and 9.2 are about 95 and 40 h, respectively. Incubation of human serum albumin (HSA, 0.5 mM) with a 10-fold molar excess of PIT-PEG (3K or 5K) at pH 6.5 and 9.2 generated PEG-HSA conjugates with average of 3.5 and 6.0 PEG chains per HSA molecule, respectively. The circular dichroism spectra of the conjugates showed that PEGylation of HSA has little influence on the secondary structure of HSA. The hexaPEGylated HSA, (TCP-PEG5K)6-HSA, exhibited very high hydrodynamic volume, and the molecular radius of HSA increased from 3.95 to 6.57 nm on hexaPEGylation. The hexaPEGylation also increased the viscosity of 4% HSA from 1.05 to 2.10 cP, and the colloid osmotic pressure from 15.2 to 48.0 mmHg. The large increase in the hydrodynamic volume and the solution properties of (TCP-PEG5K)6-HSA suggest that it could be a potential candidate as a plasma volume expander. PIT-PEG is a useful addition to the spectrum of functionalized PEG reagents available for surface decoration of proteins with PEG.

Original languageEnglish (US)
Pages (from-to)1352-1360
Number of pages9
JournalBioconjugate Chemistry
Volume19
Issue number7
DOIs
StatePublished - Jul 2008

Fingerprint

Serum Albumin
Polyethylene glycols
Hydrodynamics
Proteins
Hydrolysis
Plasma Substitutes
Cross-Linking Reagents
Osmotic Pressure
Colloids
Circular Dichroism
Viscosity
Membrane Proteins
Phosphates
Costs and Cost Analysis
phenylisothiocyanate
Dichroism
Functional groups
Condensation
Nuclear magnetic resonance
Plasmas

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

PEGylation of human serum albumin : Reaction of PEG-phenyl-isothiocyanate with protein. / Meng, Fantao; Manjula, Belur N.; Smith, Paul K.; Acharya, Seetharama A.

In: Bioconjugate Chemistry, Vol. 19, No. 7, 07.2008, p. 1352-1360.

Research output: Contribution to journalArticle

Meng, Fantao ; Manjula, Belur N. ; Smith, Paul K. ; Acharya, Seetharama A. / PEGylation of human serum albumin : Reaction of PEG-phenyl-isothiocyanate with protein. In: Bioconjugate Chemistry. 2008 ; Vol. 19, No. 7. pp. 1352-1360.
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