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  • Synthesis of semitelechelic maleimide poly(PEGA) for protein conjugation by RAFT polymerization.

Synthesis of semitelechelic maleimide poly(PEGA) for protein conjugation by RAFT polymerization.

Biomacromolecules (2009-06-10)
Emmanuelle Bays, Lei Tao, Chien-Wen Chang, Heather D Maynard
ABSTRACT

Maleimide end functionalized polymers for site-selective conjugation to free cysteines of proteins were synthesized using reversible addition-fragmentation chain transfer (RAFT) polymerization. A furan-protected maleimide chain transfer agent (CTA) was employed in the RAFT polymerization of poly(ethylene glycol) methyl ether acrylate (PEGA). Gel permeation chromatography (GPC) with laser light scattering detection indicated that 20,000 and 39,000 Da polyPEGA had been made with polydispersity indices of 1.25 and 1.36, respectively. The maleimide group on the polymer chain end was exposed by heating the poly(PEGA)s for 4 h. The deprotection efficiency was estimated to be 80 and 60% for poly(PEGA)(20 kDa) and poly(PEGA)(39 kDa), respectively. Maleimide-poly(PEGA)s were conjugated to V131C T4 lysozyme (T4L), and the resultant polymer-protein conjugates were characterized by size exclusion chromatography and gel electrophoresis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(ethylene glycol) bis(amine), Mw 10,000
Sigma-Aldrich
Poly(ethylene glycol) bis(amine), Mw 6,000
Sigma-Aldrich
Poly(ethylene glycol) bis(amine), Mw 2,000
Sigma-Aldrich
Poly(ethylene glycol) bis(amine), Mw 3,000
Sigma-Aldrich
Poly(ethylene glycol) bis(amine), Mw 20,000
Sigma-Aldrich
Poly(ethylene glycol) bis(amine), average Mn 3400
Sigma-Aldrich
Poly(ethylene glycol) diamine, average Mn 2,000
Sigma-Aldrich
Poly(ethylene glycol) diamine, average Mn 3,000
Sigma-Aldrich
Poly(ethylene glycol) diamine, average Mn 10,000
Sigma-Aldrich
Poly(ethylene glycol) diamine, average Mn 6,000