Short Communication 
Large-Scale Purification of Human Apolipoprotein Kringle Domain V (rHualkV) Expressed in Pichia pastoris
Biotechnology, 2015, 14(4), 206-210.
Abstract
Human apolipoprotein kringle domain V (rHualkV) that has an anti-angiogenic activity was expressed and purified in large scale from Pichia pastoris. rHualkV was captured by SP-Streamline resin directly from 40 L culture broth through batch type adsorption with about 80% yield (n = 8). Captured rHualkV was further purified by SP-Sepharose column chromatography and concentrated by membrane filtration. On average 5 g of rHualkV was purified per batch with the 56% final yield. The result of analytical High-Performance Liquid Chromatography (HPLC) of the purified rHualkV showed the single peak. ESI mass-spectrometry analysis of the rHualkV has proved that the purified rHualkV has the 9676 kDa that is the correct molecular weight as the one calculated by amino acid sequence. In addition, rHualkV has the same N and C-terminal residue as the expected one from DNA sequence of rHualkV gene in Pichia pastoris. Thus, through three steps of process, the anti-angiogenic rHualkV was successfully expressed and purified from Pichia pastoris, which will be able to provide a basis for much larger expression and purification for production needed for clinical evaluation later.
ASCI-ID: 11-810
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