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High-yield Purification and Characterization of Recombinant Human Leukotactin-1 in Pichia pastoris  

Lim, In-Hwan (Mogam Biotechnology Institute)
Lee, Kong-Ju (Mogam Biotechnology Institute)
Lee, Eun-Kyoung (Mogam Biotechnology Institute)
Park, Mu-Rim (Mogam Biotechnology Institute)
Lee, Gue-Wha (Mogam Biotechnology Institute)
Yeup Yoon (Mogam Biotechnology Institute)
Park, Doo-Hong (Mogam Biotechnology Institute)
Jung, Kyung-Hwan (Mogam Biotechnology Institute, Department of Food and Biotechnology, Chungju National University)
Publication Information
Biotechnology and Bioprocess Engineering:BBE / v.9, no.1, 2004 , pp. 1-6 More about this Journal
Abstract
The human chemokine, the short version of leukotactin-1(shLkn-1;molecular weight=7.2 kD and 66 amino acids), was expressed and secreted into a culture medium using the me-thylotrophic yeast, Pichia pastoris. The recombinant shLkn-1 was purified from the culture supernatant using a simple two-step procedure consisting of cation exchange and reverse phase chromatography(RPC), in which shLkn-1 was highly purified (99.5%) with a high recovery yield of 82.7%. The C-terminal truncated derivative of shLkn-1 was found in the supernatant and was separated by RPC. The physicochemical properties of the purified shLkn-1 were verified to be the same as expected. The biological activity of the purified recombinant shLkn-1 was also quantified using a chemotaxis assay. It was observed that the recombinant shLkn-1 had the maximum migration activity at a concentration of 10nM, as potent as MIP-1${\alpha}$.
Keywords
C-terminal truncation; leukotactin; Pichia pastoris; secretion;
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