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Title: Purification of recombinant antigen <i style="">Bm</i>SXP used in panLF Rapid kit for lymphatic filariasis
Authors: Khoo, Teng Kew
Noordin, Rahmah
Santhanam, Amutha
Keywords: BmSXP Recombinant Antigen
Lymphatic Filariasis
PanLF Rapid
Purification Optimizations
Issue Date: Apr-2012
Publisher: NISCAIR-CSIR, India
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 <i>B. malayi </i>antigens, <i>Bm</i>R1 and <i style="">Bm</i>SXP. 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 <i style="">Bm</i>SXP 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 m<i style="">M</i> imidazole concentration in lysis buffer, 30 m<i style="">M</i> imidazole concentration in wash buffer, and 10 column volume wash buffer containing 300 m<i style="">M</i> 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 <i style="">Bm</i>SXP 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.
Description: 256-264
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.50(04) [April 2012]

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