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

Martina Tuttolomondo; Pernille Lund Hansen; Jan Mollenhauer; Henrik J Ditzel

doi:10.1016/j.ab.2017.11.015

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

Anal Biochem. 2018 Feb 1:542:16-19. doi: 10.1016/j.ab.2017.11.015. Epub 2017 Nov 21.

Authors

Martina Tuttolomondo¹, Pernille Lund Hansen², Jan Mollenhauer², Henrik J Ditzel³

Affiliations

¹ Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, Odense, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark. Electronic address: mtuttolomondo@health.sdu.dk.
² Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, Odense, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
³ Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, Odense, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark; Department of Oncology, Odense University Hospital, Odense, Denmark. Electronic address: hditzel@health.sdu.dk.

PMID: 29169777
DOI: 10.1016/j.ab.2017.11.015

Abstract

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.

Keywords: DMBT1; PRM; Pattern recognition receptor; Protein purification; Salivary agglutinin (SAG); gp340.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium-Binding Proteins
Cell Line, Tumor
Chromatography, Gel
Chromatography, Liquid
DNA-Binding Proteins
Humans
Receptors, Cell Surface / chemistry
Receptors, Cell Surface / isolation & purification*
Receptors, Cell Surface / metabolism*
Recombinant Proteins / chemistry
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Streptococcus mutans / chemistry
Streptococcus mutans / metabolism
Tumor Suppressor Proteins

Substances

Calcium-Binding Proteins
DMBT1 protein, human
DNA-Binding Proteins
Receptors, Cell Surface
Recombinant Proteins
Tumor Suppressor Proteins