Preparation of a novel polymer monolith with functional polymer brushes by two‐step atom‐transfer radical polymerization for trypsin immobilization

Abstract

Novel porous polymer monoliths grafted with poly{oligo[(ethylene glycol) methacrylate]-co-glycidyl methacrylate} brushes were fabricated via two-step atom-transfer radical polymerization and used as a trypsin-based reactor in a continuous flow system. This is the first time that atom-transfer radical polymerization technique was utilized to design and construct polymer monolith bioreactor. The prepared monoliths possessed excellent permeability, providing fast mass transfer for enzymatic reaction. More importantly, surface properties, which were modulated via surface-initiated atom-transfer radical polymerization, were found to have a great effect on bioreactor activities based on Michaelis-Menten studies. Furthermore, three model proteins were digested by the monolith bioreactor to a larger degree within dramatically reduced time (50 s), about 900 times faster than that by free trypsin (12 h). The proposed method provided a platform to prepare porous monoliths with desired surface properties for immobilizing various enzymes.

Tunable Polymeric Scaffolds for Enzyme Immobilization

“…However, the main drawback of polymer monoliths relies on the large size of their pores, which results in low enzyme loads. Fortunately, this limitation can be circumvented by grafting a polymer layer on the surface of the macropores of monoliths (Peterson et al, 2003;Li et al, 2014;Wen et al, 2016b). Enzyme-monolith hybrids ( Figure 9A) have a relevant impact in microfluidic applications (Logan et al, 2007;Krenkova et al, 2009;Meller et al, 2017;Cheng et al, 2019) and, in less extension, biosensors (Luo et al, 2019), and HPLC (Girelli and Mattei, 2005).…”

“…However, the main drawback of polymer monoliths relies on the large size of their pores, which results in low enzyme loads. Fortunately, this limitation can be circumvented by grafting a polymer layer on the surface of the macropores of monoliths ( Peterson et al, 2003 ; Li et al, 2014 ; Wen et al, 2016b ).…”

Reversible Two‐Enzyme Coimmobilization on pH‐Responsive Imprinted Monolith for Glucose Detection

“…The drawback of porous polymer monoliths relies on their small surface areas due to the absence of small pores, which might result in low immobilization capacities for enzymes. This limitation can be circumvented by grafting a polymer layer on the surface of macropores of monoliths …”