Macroporous Polymer–Protein Hybrid Materials for Antibody Purification by Combination of Reactive Gelation and Click-Chemistry

• Main Authors: Marcel Lorenz, Carolina Paganini, Giuseppe Storti, Massimo Morbidelli
• Format: Article
• Language: English
• Published: MDPI AG 2019-05-01
• Series: Materials
• Subjects: macroporous Materials; click-chemistry; reactive gelation; protein immobilization; antibody purification
• Online Access: https://www.mdpi.com/1996-1944/12/10/1580

Clickable core-shell nanoparticles based on poly(styrene-<i>co-</i>divinylbenzene-<i>co-</i>vinylbenzylazide) have been synthesized via emulsion polymerization. The 38 nm sized particles have been swollen by divinyl benzene (DVB) and 2,2&#8217;-azobis(2-methylpropionitrile) (AIBN) and subsequently processed under high shear rates in a Z-shaped microchannel giving macroporous microclusters (100 &#181;m), through the reactive gelation process. The obtained clusters were post-functionalized by &#8220;click-chemistry&#8222; with propargyl-PEG-NHS-ester and propargylglicidyl ether, yielding epoxide or NHS-ester activated polymer supports for bioconjugation. Macroporous affinity materials for antibody capturing were produced by immobilizing recombinant <i>Staphylococcus aureus</i> protein A on the polymeric support. Coupling chemistry exploiting thiol-epoxide ring-opening reactions with cysteine-containing protein A revealed up to three times higher binding capacities compared to the protein without cysteine. Despite the lower binding capacities compared to commercial affinity phases, the produced polymer&#8211;protein hybrids can serve as stationary phases for immunoglobulin affinity chromatography as the materials revealed superior intra-particle mass transports.