bc9b00839_si_002.pdf (766.37 kB)
Site Directed Disulfide PEGylation of Interferon-β-1b with Fork Peptide Linker
journal contribution
posted on 2020-03-04, 22:44 authored by Shayan Abbasi, Homa Farahani, Hossein Lanjanian, Mohammad Taheri, Loghman Firoozpour, Jamshid Davoodi, Sama Pirkalkhoran, GholamHossein Riazi, Shahriar PooyanThe attachment of
PEG to biopharmaceuticals has been applied for
enhancement of bioavailability and improved stability. The PEG polymer
is highly hydrated; thus effective attachment to inaccessible sites
could be hindered. We have devised a scheme to address this issue
by introducing a considerable distance between PEG and protein by
addition of a linear peptide, appended to long chained reactive linkers.
Second, the position of PEG conjugation directly affects biological
activity. Accordingly, a disulfide bond could be considered as an
ideal choice for site directed PEGylation; but reactivity of both
thiol moieties to bridging reagent is critical for maintenance of
protein structure. In our design, a forked structure with two arms
provides essential flexibility to account for dissociation of reduced
cysteines. An efficient yield for disulfide PEGylation of IFN-β1b
was attained and specificity, biophysical characterization, biological
activity, and pharmacokinetics were surveyed.