One-step FPLC-size-exclusion chromatography procedure for purification of rDMBT1 6 kb with increased biological activity

Deleted in Malignant Brain Tumor 1 (DMBT1, alias SAG or gp340) is a pattern recognition receptor involved in immune defense, cell polarization, differentiation and regeneration. To investigate the role of the protein in physiological and pathological processes, the protein has often been isolated fr...

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Bibliographic Details
Published in:Analytical biochemistry 2018-02, Vol.542, p.16-19
Main Authors: Tuttolomondo, Martina, Hansen, Pernille Lund, Mollenhauer, Jan, Ditzel, Henrik J.
Format: Article
Language:English
Subjects:
DMBT1
gp340
Pattern recognition receptor
PRM
Protein purification
Salivary agglutinin (SAG)
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Summary:Deleted in Malignant Brain Tumor 1 (DMBT1, alias SAG or gp340) is a pattern recognition receptor involved in immune defense, cell polarization, differentiation and regeneration. To investigate the role of the protein in physiological and pathological processes, the protein has often been isolated from saliva or produced in vitro and purified by a multistep affinity purification procedure using bacteria, followed by FPLC. Here, we compared a simple, one-step FPLC-SEC protocol for purification of recombinant DMBT1 6 kb, with that of the standard bacteria affinity purification-based protocol. Our data suggest that our FPLC-SEC protocol yields DMBT1 in a more native conformation.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2017.11.015