Production and purification of recombinant Blomia tropicalis group 5 allergen from Pichia pastoris culture |
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| Authors: | Lin-Tang Goh I-Chun Kuo Sa Luo Kaw Yan Chua Moshe White |
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| Institution: | (1) Bioprocessing Technology Centre, Singapore;(2) Department of Paediatrics, The National University of Singapore, 10 Kent Ridge, Crescent, Singapore, 119260 |
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| Abstract: | The gene of Blomia tropicalis group 5 allergen (Blo t 5) was cloned and expressed in Pichia pastoris KM71. Selected KM71 clones were cultivated in a fed-batch bioreactor feeding first glycerol then followed by methanol. Recombinant Blo t 5 constituted about 30% of the total broth protein after 60 h cultivation. The harvested broth was purified to >95% purity by a two-step anion exchange chromatography. The overall yield was 37 mg Blo t 5 per litre broth. |
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| Keywords: | anion exchange chromatography Blomia tropicalis dust mite allergen fed-batch culture Pichia pastoris |
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