A versatile method for the conjugation of proteins and peptides to poly[2-(dimethylamino)ethyl methacrylate]

van Dijk-Wolthuis, W.N.E., van de Wetering, P., Hinrichs, W.L.J., Hofmeyer, L.J.F., Liskamp, R.M.J. , Crommelin, D.J.A. and Hennink, W.E. (1999) A versatile method for the conjugation of proteins and peptides to poly[2-(dimethylamino)ethyl methacrylate]. Bioconjugate Chemistry, 10(4), pp. 687-692. (doi: 10.1021/bc980126+)

Abstract

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-NH2 was coupled to the thiolated polymer via a thioether linkage. The coupling efficiency, as determined by GPC (Tf), SDS−PAGE [F(ab‘)], or 1H 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.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Liskamp, Professor Robert
• Authors: van Dijk-Wolthuis, W.N.E., van de Wetering, P., Hinrichs, W.L.J., Hofmeyer, L.J.F., Liskamp, R.M.J., Crommelin, D.J.A., and Hennink, W.E.
• College/School: College of Science and Engineering > School of Chemistry
• Journal Name: Bioconjugate Chemistry
• ISSN: 1043-1802
• ISSN (Online): 1520-4812