posted on 2021-06-08, 19:12authored byValentina
J. Liebich, Olga Avrutina, Jan Habermann, Laura M. Hillscher, Markus Langhans, Tobias Meckel, Markus Biesalski, Harald Kolmar
Herein, we report a novel two-step
method for the covalent, site-directed,
and efficient immobilization of proteins on lab-made paper sheets.
First, paper fibers were modified with a peptidic anchor comprising
enzyme recognition motifs. Four different conjugation strategies for
peptide immobilization were evaluated with respect to reproducibility
and fiber loading efficiency. After manufacturing of the peptide-preconditioned
paper, oriented conjugation of the model protein tGFP containing a
C-terminal recognition sequence for either sortase A or microbial
transglutaminase was assessed semiquantitatively by fluorescence measurement
and inspected by confocal laser scanning microscopy (CLSM). The two
enzymes utilized for protein conjugation used the same oligoglycine
peptide anchor, and both proved to be suitable for controlled oriented
linkage of substrate proteins at physiological conditions.