Crystallization of antiangiogenic Kringle V derived from human apolipoprotein A: Crystallization applied to purification and formulation

Kwan Yub Kang, Jung Hwan Park, In Hwan Lim, Sung Geun Kim, See Hyoung Park, Won Kyung Kim, Jeong Woon Hong, Hyun Kyung You, Kyung Hwan Jung, Chan Wha Kim

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7 Citations (Scopus)

Abstract

In this study, the Kringle V domain (Glu4225-Ser4310) of human apolipoprotein A, an antiangiogenic polypeptide, was expressed as a secreted form in Pichia pastoris, and was purified via a process consisting of three chromatographic steps. The chromatographically purified kringle V domain contained a C-terminal serine-deleted form and several high-molecular-weight forms, which were suspected to represent glycosylated derivatives. In order to remove these derivatives, we employed a crystallization process. The crystallization of kringle V resulted in an 85% recovery yield, and also resulted in the complete removal of the aforementioned high-molecular-weight forms. However, we were still able to detect a trace of the C-terminal serine-deleted form. The prepared Kringle V crystals were stable within a pH range of 7.0 to 8.0, and were completely dissolved by dilution, which is a crucial factor in the preparation of a highly concentrated formulation. The chromatogram of the crystallized kringle V on reversed-phase HPLC analysis was identical to that observed without crystallization. Also, we noted that the original anti-wound migration activities of the molecule toward human umbilical vein endothelial cells were completely retained.

Original languageEnglish
Pages (from-to)916-925
Number of pages10
JournalBioscience, Biotechnology and Biochemistry
Volume70
Issue number4
DOIs
Publication statusPublished - 2006

Keywords

  • Antiangiogenic polypeptide
  • Apolipoprotein A
  • Crystal
  • Formulation
  • Kringle

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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  • Cite this

    Kang, K. Y., Park, J. H., Lim, I. H., Kim, S. G., Park, S. H., Kim, W. K., Hong, J. W., You, H. K., Jung, K. H., & Kim, C. W. (2006). Crystallization of antiangiogenic Kringle V derived from human apolipoprotein A: Crystallization applied to purification and formulation. Bioscience, Biotechnology and Biochemistry, 70(4), 916-925. https://doi.org/10.1271/bbb.70.916