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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.50 [2012] >
IJEB Vol.50(04) [April 2012] >
| Title: | Purification of recombinant antigen BmSXP 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 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. |
| Page(s): | 256-264 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.50(04) [April 2012]
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