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Cytotechnology

, Volume 10, Issue 2, pp 137–146 | Cite as

Purification and characterization of immunoglobulin production stimulating factor-IIβ derived from Namalwa cells

  • Takuya Sugahara
  • Hiroto Nakajima
  • Sanetaka Shirahata
  • Hiroki Murakami
Original Research Papers

Abstract

Two immunoglobulin production stimulating factors (IPSF) have been found in human Burkitt's lymphoma Namalwa cells. One IPSF named IPSF-IIα was purified and identified as glyceraldehyde-3-phosphate dehydrogenase as previously reported. We report here purification, identification and characterization of IPSF-IIβ. IPSF-IIβ was purified by the serial use of ammonium sulfate fractionation, hydrophobic interaction column chromatography, anion-exchange column chromatography and gel filtration. The IPSF-IIβ was estimated as a 46 KD monomeric polypeptide by gel filtration and SDS-PAGE. Partial amino acid sequence of the 46 KD protein was analyzed for 26 amino acid residues. The sequence very closely coincided with enolase (EC 4.2.1.11) derived from various origins and, it was completely homologous with that of human enolase α-chain. Rabbit muscle enolase stimulated IgM production of hybridoma lines, and IPSF-IIβ had the enzymic activity. These results suggested that IPSF-IIβ was α-enolase or its isozyme. IPSF activities of IPSF-IIβ was stable in alkaline conditions whereas the enzymic activity was rapidly lost in alkaline conditions. Though IPSF-IIβ stimulated IgM production of both human-human and mouse-mouse hybridoma lines in serum-free condition, it partially suppressed IgE production of mouse-mouse hybridoma lines.

Key words

immunoglobulin production stimulating factor human enolase α-chain Burkitt's lymphoma cell line Namalwa cells human-human hybridoma serum-free medium 

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Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Takuya Sugahara
    • 1
  • Hiroto Nakajima
    • 1
  • Sanetaka Shirahata
    • 1
  • Hiroki Murakami
    • 1
  1. 1.Graduate School of Genetic Resources TechnologyKyushu University 46-09FukuokaJapan

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