Site-Specific DBCO Modification of DEC205 Antibody for Polymer Conjugation

Polymers

Rudolf Zentel¹  Regina Holm¹  Matthias Barz¹  Simone Beck¹  Meike Schinnerer²  Kaloian Koynov³  Jennifer Schultze³  Hans-Joachim Räder³ 

[1] Institute of Organic Chemistry, Johannes-Gutenberg University Mainz, Duesbergweg 10-14, D-55128 Mainz, Germany;Institute of Physical Chemistry, Jakob Welder Weg 11, D-55128 Mainz, Germany;Max-Planck-Institute for Polymer Research, Ackermannweg 10, D-55128 Mainz, Germany;

关键词: cancer immune therapy;    vaccination;    dendritic cells (DCs);    pDNA polyplex;    RAFT polymerization;    targeting;    DEC205 antibody;    bioorthogonal chemistry;    strain-promoted alkyne-azide cycloaddition (SPAAC);   

DOI  :  10.3390/polym10020141

来源: DOAJ

【 摘 要 】

The design of multifunctional polymer-based vectors, forming pDNA vaccines, offers great potential in cancer immune therapy. The transfection of dendritic immune cells (DCs) with tumour antigen-encoding pDNA leads to an activation of the immune system to combat tumour cells. In this work, we investigated the chemical attachment of DEC205 antibodies (aDEC205) as DC-targeting structures to polyplexes of P(Lys)-b-P(HPMA). The conjugation of a synthetic block copolymer and a biomacromolecule with various functionalities (aDEC205) requires bioorthogonal techniques to avoid side reactions. Click chemistry and in particular the strain-promoted alkyne-azide cycloaddition (SPAAC) can provide the required bioorthogonality. With regard to a SPAAC of both components, we firstly synthesized two different azide-containing block copolymers, P(Lys)-b-P(HPMA)-N3(stat) and P(Lys)-b-P(HPMA)-N3(end), for pDNA complexation. In addition, the site-specific incorporation of ring-strained dibenzocyclooctyne (DBCO) moieties to the DEC205 antibody was achieved by an enzymatic strategy using bacterial transglutaminase (BTG). The chemical accessibility of DBCO molecules within aDEC205 as well as the accessibility of azide-functionalities on the polyplex’ surface were investigated by various SPAAC experiments and characterized by fluorescence correlation spectroscopy (FCS).

【 授权许可】

Unknown