title: Purification of recombinant antigen BmSXP used in panLF rapid kit for lymphatic filariasis.
abstract: A rapid antibody detection test is very useful for the detection of lymphatic filariasis, especially for certification and surveillance of post-mass drug administration. panLF Rapid kit is suitable for this purpose since it can detect all species of lymphatic filaria. It is based on the detection of anti-filarial IgG4 antibodies that react with recombinant B. malayi antigens, BmR1 and BmSXP. There is an increase demand for the test due to its attributes of being rapid, sensitive and specific results, as well as its field-applicability. The main aim of this paper is to obtain high recovery and purity of recombinant antigen BmSXP via a modified method of immobilized metal affinity chromatography (IMAC). The highest product yield of 11.82 mg/g dry cell weight (DCW) was obtained when IMAC was performed using the optimized protocol of 10 mM imidazole concentration in lysis buffer, 30 mM imidazole concentration in wash buffer, and 10 column volume wash buffer containing 300 mM salt concentration. This gave a 54% protein recovery improvement over the manufacturer's protocol which recorded a product yield of only 7.68 mg/g DCW. The recovered BmSXP recombinant antigen showed good western blot reactivity, high sensitivity (31/32, 97%) and specificity (32/32, 100%) in ELISA, thus attesting to its good purity and quality.
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia (USM), 11800 Pulau Pinang, Malaysia.
authors.forename: Teng Kew
authors.initials: TK
authors.lastname: Khoo
authors.forename: Rahmah
authors.initials: R
authors.lastname: Noordin
authors.forename: Amutha
authors.initials: A
authors.lastname: Santhanam
mesh_terms: D000947:Antigens, Helminth / Q000032:analysis* / Q000302:isolation & purification; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004605:Elephantiasis, Filarial / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification
chemical_list: D000947:Antigens, Helminth; D011994:Recombinant Proteins