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Production and purification of recombinant Blomia tropicalis group 5 allergen from Pichia pastoris culture

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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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Authors and Affiliations

  1. Bioprocessing Technology Centre, Singapore

    Lin-Tang Goh, Sa Luo & Moshe White

  2. Department of Paediatrics, The National University of Singapore, 10 Kent Ridge, Crescent, Singapore, 119260

    I-Chun Kuo & Kaw Yan Chua

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  1. Lin-Tang Goh
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  2. I-Chun Kuo
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  3. Sa Luo
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  4. Kaw Yan Chua
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  5. Moshe White
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Goh, LT., Kuo, IC., Luo, S. et al. Production and purification of recombinant Blomia tropicalis group 5 allergen from Pichia pastoris culture. Biotechnology Letters 23, 661–665 (2001). https://doi.org/10.1023/A:1010323611039

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  • DOI: https://doi.org/10.1023/A:1010323611039

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