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High-yield purification and characterization of recombinant human leukotactin-1 inPichia pastoris

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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 methylotrophic 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 10 nM, as potent as MIP-1α.

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Authors and Affiliations

  1. Mogam Biotechnology Institute, 341 Pojung-Ri, Koosung-Eup, Yougin, 449-913, Kyonggi-Do, Korea

    In Hwan Lim, Kong Ju Lee, Eun Kyoung Lee, Mu Rim Choi, Gue-Wha Lee, Yeup Yoon, Doo-Hong Park & Kyung-Hwan Jung

  2. Department of Food and Biotechnology, Chungju National University, 380-702, Chungbuk, Korea

    Kyung-Hwan Jung

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  1. In Hwan Lim
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  2. Kong Ju Lee
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  3. Eun Kyoung Lee
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  5. Gue-Wha Lee
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Correspondence to Kyung-Hwan Jung.

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Lim, I.H., Lee, K.J., Lee, E.K. et al. High-yield purification and characterization of recombinant human leukotactin-1 inPichia pastoris . Biotechnol Bioproc E 9, 1–6 (2004). https://doi.org/10.1007/BF02949314

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  • DOI: https://doi.org/10.1007/BF02949314

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