Abstract
We developed a spongy-like porous polymer (spongy monolith) consisting of poly(ethylene-co-glycidyl methacrylate) with continuous macropores that allowed efficient in situ reaction between the epoxy groups and proteins of interest. Immobilization of protein A on the spongy monolith enabled high-yield collection of immunoglobulin G (IgG) from cell culture supernatant even at a high flow rate. In addition, immobilization of pepsin on the spongy monolith enabled efficient online digestion at a high flow rate.
Highlights
We developed a spongy-like porous polymer consisting of poly(ethylene-co-glycidyl methacrylate) with continuous macropores that allowed efficient in situ reaction between the epoxy groups and proteins of interest
We prepared a novel spongy monolith consisting of poly(ethylene-co-glycidyl methacrylate) (PEGM)
After the monolith was packed into a column, protein A was immobilized onto the media in situ, and the affinity reaction was quantitatively examined and validated under high-throughput conditions
Summary
We developed a spongy-like porous polymer (spongy monolith) consisting of poly(ethylene-co-glycidyl methacrylate) with continuous macropores that allowed efficient in situ reaction between the epoxy groups and proteins of interest. Immobilization of protein A on the spongy monolith enabled high-yield collection of immunoglobulin G (IgG) from cell culture supernatant even at a high flow rate. To obtain a high-quality antibody medicine at low cost, it is necessary to select highly productive cells, optimize the culture conditions, and develop an efficient purification method. To evaluate the productivity of a system for biosynthesis of an antibody, especially of the immunoglobulin G (IgG) subtype, a chromatographic system using a protein A immobilized column is often employed for selection and optimization of the cell culture. The control of pore size, especially for larger pore (>10 μm), scale up in column size, and packing to columnar tubes are not easy Instead of these typical monoliths, we proposed using a sponge-like material or spongy monolith as a novel separation medium[32, 33]. We expected that the spongy monolith containing specific functional groups, such as epoxy groups, would be useful for affinity chromatography and overcome the limitations of current media
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