A versatile method for the conjugation of proteins and peptides to poly[2-(dimethylamino)ethyl methacrylate].
Random copolymers of 2-(dimethylamino) ethyl methacrylate (DMAEMA) with aminoethyl methacrylate (AEMA) were synthesized by radical polymerization. The amount of incorporated primary amino groups could be controlled by the feed ratio of AEMA to DMAEMA, and was varied from 2 to 6 mol %. Subsequently, protected thiol groups were introduced in a derivatization step with N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and subsequent treatment with dithiothreitol (DTT). The obtained thiolated p(DMAEMA-co-AEMA) was conjugated to transferrin (Tf) or the F(ab') fragment of mAb 323/A3 via a disulfide linkage. Moreover, the maleimide derivative of the nuclear localization signal (NLS) decapeptide Gly-Pro-Lys-Lys-Lys-Arg-Lys-Val-Glu-Asp-NH(2) was coupled to the thiolated polymer via a thioether linkage. The coupling efficiency, as determined by GPC (Tf), SDS-PAGE [F(ab')], or (1)H NMR (NLS peptide) was 90-95% for the Tf conjugate, and more than 95% for the F(ab') conjugate and the NLS conjugate. The synthetic strategy described in this paper is a universal method for the preparation of conjugates of proteins and peptides with pDMAEMA in particular. This method can possibly be used to synthesize protein-polymethacrylate conjugates in general.
van Dijk-Wolthuis
W N
WN
Department of Pharmaceutics, Sorbonnelaan 16, Department of Medicinal Chemistry, Utrecht Institute for Pharmaceutical Sciences (UIPS), Faculty of Pharmacy, Utrecht University, PO Box 80 082, 3508 TB Utrecht, The Netherlands.
van de Wetering
P
P
Hinrichs
W L
WL
Hofmeyer
L J
LJ
Liskamp
R M
RM
Crommelin
D J
DJ
Hennink
W E
WE
D000911:Antibodies, Monoclonal / Q000737:chemistry; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008768:Methylmethacrylates / Q000737:chemistry*; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013439:Sulfhydryl Reagents; D014168:Transferrin / Q000737:chemistry; D014783:Viscosity
D000911:Antibodies, Monoclonal; D000952:Antigens, Polyomavirus Transforming; D007140:Immunoglobulin Fab Fragments; D008768:Methylmethacrylates; D010455:Peptides; D011506:Proteins; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D014168:Transferrin; C035600:dimethylaminoethyl methacrylate-methylmethacrylate copolymer