title: Purification of BmR1 recombinant protein.
abstract: This paper describes a refinement in the purification step that facilitated the downstream recovery of high purity BmR1 recombinant protein, which is a protein used as a test reagent in the commercialized rapid tests for detection of lymphac filariasis i.e. Brugia Rapid™ and panLF rapid™. Purification was performed by immobilized metal affinity chromatography (IMAC), followed by ion exchange chromatography (IEX). Results showed that a total of 10.27 mg of BmR1 was obtained when IMAC was performed using 20 mM of imidazole and 5 column volume of wash buffer containing 500 mM of NaCl. Purity of the target protein was enhanced when buffer at pH 5.8 was used during the IEX. Two proteins that recurrently appeared below the BmR1 recombinant protein were identified by mass-spectrometry analysis as the same protein, thus they were probably degradation products of BmR1. These strategies improve purity of the target protein to be used in applications such as production of aptamers and monoclonal antibodies.
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, Penang, Malaysia.
authors.forename: Norsyahida
authors.initials: N
authors.lastname: Arifin
authors.forename: Madihah
authors.initials: M
authors.lastname: Basuni
authors.forename: Chew Ai
authors.initials: CA
authors.lastname: Lan
authors.forename: Ahmad Ramli Mohd
authors.initials: AR
authors.lastname: Yahya
authors.forename: Rahmah
authors.initials: R
authors.lastname: Noordin
mesh_terms: D000818:Animals; D017178:Brugia malayi / Q000235:genetics / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D015801:Helminth Proteins; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins