Optimization of the expression of surface antigen SAG1/2 of Toxoplasma gondii in the yeast Pichia pastoris

Thiruvengadam, G. and Ithoi, Init and Fong, Mun Yik and Lau, Yee Ling (2011) Optimization of the expression of surface antigen SAG1/2 of Toxoplasma gondii in the yeast Pichia pastoris. Tropical Biomedicine, 28 (3). pp. 506-513. ISSN 0127-5720,

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Abstract

Surface antigens are the most abundant proteins found on the surface of the parasite Toxoplasma gondii. Surface antigen 1 (SAG1) and Surface antigen 2 (SAG2) remain the most important and extensively studied surface proteins. These antigens have been identified to play a role in host cell invasion, immune modulation, virulence attenuation. Recombinant SAG1/2 was cloned and expressed in yeast Pichia pastoris. We describe here optimization of critical parameters involved in high yield expression of the recombinant SAG1/2. Our results suggest that recombinant SAG1/2 were best expressed at 30ºC, pH 6 and 1 methanol as the carbon source by X33 Pichia cells. Additional optimizations included the downstream process such as ammonium sulphate precipitation and dialysis. The fusion protein was purified using Ni-NTA purification system with 80 recovery. The purified protein was 100 specific and sensitive in detection of toxoplasmosis.

Item Type:	Article
Funders:	UNSPECIFIED
Additional Information:	Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia * Corresponding author email: lauyeeling@um.edu.my
Uncontrolled Keywords:	Parasitology; antigen SAG1/2; Toxoplasma gondii; yeast Pichia pastoris
Subjects:	R Medicine
Divisions:	Faculty of Medicine
Depositing User:	Fong Mun Yik
Date Deposited:	07 Jun 2012 07:06
Last Modified:	14 Nov 2019 04:17
URI:	http://eprints.um.edu.my/id/eprint/3219

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