Production, purification and diagnostic application of filarial recombinant protein WbSXP-1 expressed in salt inducible Escherichia coli.
Wuchereria bancrofti protein WbSXP-1 was identified and established as a potential candidate for the diagnosis of lymphatic filariasis. For the economic production of rWbSXP-1, osmotically (salt) inducible Escherichia coli GJ1158 was preferred. Cultivation and expression was optimized in 3 L airlift bioreactor (ALB) and was successfully extended to 30 L ALB. Purification of rWbSXP-1 his-tag protein was optimized in technical scale using FPLC and the maximal recovery of rWbSXP-1 with significant level of purity was achieved using the combination of IMAC and gel filtration. Quality criteria for immuno-reactivity of purified rWbSXP-1 were established for diagnostic applications. Enhancement of sensitivity in rapid diagnostic format was optimized to effectively detect weak to strong antibody reactivity in individuals exposed to lymphatic filariasis. Performance of the rapid format during field evaluation was successful. The accelerated stability assessment of the rapid format satisfied the requirements of WHO-cGMP norms. This investigation presents a successful technical scale production and purification of rWbSXP-1 considering the future industrial application and an enhanced rapid flow through antibody assay for the diagnosis of human lymphatic filariasis.
Centre for Biotechnology, Anna University, Chennai, India.
S
S
Janardhan
P
P
Pandiaraja
S
S
Thirugnanam
M N
MN
Balamurali
Kennedy
K
Fernando
H C
HC
Mody
P K
PK
Desai
S
S
Meenakshisundaram
P
P
Kaliraj
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D000947:Antigens, Helminth / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D019149:Bioreactors; D004605:Elephantiasis, Filarial / Q000175:diagnosis* / Q000469:parasitology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D005374:Filtration / Q000295:instrumentation; D015801:Helminth Proteins / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012965:Sodium Chloride / Q000494:pharmacology; D014958:Wuchereria bancrofti / Q000235:genetics* / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins; D012965:Sodium Chloride