Self-Immobilization of Car9 Fusion Proteins within High Surface Area Silica Sol-Gels and Dynamic Control of Protein Release.
Protein entrapment within silica matrices during sol-gel formation is an effective way of producing biocatalysts with high load, activity retention, and minimal leaching. On the other hand, mesoporous silica materials have been favored for diffusional control of protein delivery because of their regular pore size and morphology and in spite of the drawback of requiring post-synthesis loading with cargo proteins. Here, we describe a hybrid technology in which fusion of the silica-binding Car9 dodecapeptide to model fluorescent proteins allows for their simultaneous entrapment and surface immobilization within sol-gel monoliths that can be fabricated in air and oil phases. Spherical particles produced by injecting a mixture of silicic acid and Car9-tagged proteins in silicone oil exhibit high surface area (>400 m2/g), 15-nm-diameter mean pore size and homogeneous protein loading. Incubation in arginine-containing buffer disrupts the interaction between Car9 extensions and silica surfaces and triggers the continuous or discontinuous (on/off) release of cargo proteins with pH-tunable kinetics. This simple approach for producing hybrid silica materials that stably encapsulate and release one or more Car9-tagged proteins in a single step may prove useful for applications requiring dynamic control of protein concentration.
Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
Wenlan
W
Yang
Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
Brittney
B
Hellner
Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
François
F
Baneyx
D001120:Arginine / Q000737:chemistry; D002021:Buffers; D002384:Catalysis; D005782:Gels / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D058428:Silica Gel / Q000737:chemistry; D012822:Silicon Dioxide
D002021:Buffers; D005782:Gels; D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D058428:Silica Gel; D012822:Silicon Dioxide; D001120:Arginine