Fine‐tuning sortase‐mediated immobilization of protein layers on surfaces using sequential deprotection and coupling |
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Authors: | Maryam Raeeszadeh‐Sarmazdeh Ranganath Parthasarathy Eric T Boder |
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Institution: | 1. Dept. of Chemical and Biomolecular Engineering, University of Tennessee, Knoxville, TN;2. Dept. of Bioengineering, University of Pennsylvania, Philadelphia, PA |
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Abstract: | Increasing interest in protein immobilization on surfaces has heightened the need for techniques enabling layer‐by‐layer protein attachment. Here, we report a technique for controlling enzyme‐mediated immobilization of layers of protein on the surface using a genetically encoded protecting group. An enterokinase‐cleavable peptide sequence was inserted at the N‐terminus of bifunctional fluorescent proteins containing Sortase A substrate recognition tags at both ends to control Sortase A‐mediated protein immobilization on the surface layer‐by‐layer. Efficient, sequential immobilization of a second layer of protein using Sortase A required removal of the N‐terminal protecting group, suggesting the method enables multilayer synthesis using cyclic deprotection and coupling steps. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:824–831, 2017 |
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Keywords: | sortase A protein immobilization protein ligation |
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