PREPARATION AND PURIFICATION OF 3-FLUORO-TYROSINE LABELED TUMOR-NECROSIS-FACTOR-ALPHA AND PRELIMINARY F-19 N.M.R. INVESTIGATION

  • Authors:
    • JK SAGOO
    • LC CHAPLIN
    • NRA BEELEY
    • C IAONNOU
    • AM NESBITT
    • SJB TENDLER
  • View Affiliations

  • Published online on: October 1, 1994     https://doi.org/10.3892/ijo.5.4.937
  • Pages: 937-943
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Abstract

A one-step protocol for the purification of recombinant human tumour necrosis factor-alpha (TNF alpha) has been developed based on the use of antibody affinity chromatography. The method allows for the preparation of large amounts of the protein (>15 mg). The overall recovery of the purified material from Esherichia coli lysate after buffer exchange into 0.8% mannitol is 48%, with no apparent loss of bioactivity. This method has been utilized for the preparation of 3-fluoro-tyrosine labelled human TNF alpha. Data indicate that the protein produced in minimal media is a heterotrimer consisting of two 17 kDa monomers and one proteolytically cleaved 14 kDa unit. Preliminary F-19 n.m.r. spectroscopy indicated that the 3-fluoro-tyrosine labelled protein is suitable for further study using this technique.

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October 1994
Volume 5 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

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APA
SAGOO, J., CHAPLIN, L., BEELEY, N., IAONNOU, C., NESBITT, A., & TENDLER, S. (1994). PREPARATION AND PURIFICATION OF 3-FLUORO-TYROSINE LABELED TUMOR-NECROSIS-FACTOR-ALPHA AND PRELIMINARY F-19 N.M.R. INVESTIGATION. International Journal of Oncology, 5, 937-943. https://doi.org/10.3892/ijo.5.4.937
MLA
SAGOO, J., CHAPLIN, L., BEELEY, N., IAONNOU, C., NESBITT, A., TENDLER, S."PREPARATION AND PURIFICATION OF 3-FLUORO-TYROSINE LABELED TUMOR-NECROSIS-FACTOR-ALPHA AND PRELIMINARY F-19 N.M.R. INVESTIGATION". International Journal of Oncology 5.4 (1994): 937-943.
Chicago
SAGOO, J., CHAPLIN, L., BEELEY, N., IAONNOU, C., NESBITT, A., TENDLER, S."PREPARATION AND PURIFICATION OF 3-FLUORO-TYROSINE LABELED TUMOR-NECROSIS-FACTOR-ALPHA AND PRELIMINARY F-19 N.M.R. INVESTIGATION". International Journal of Oncology 5, no. 4 (1994): 937-943. https://doi.org/10.3892/ijo.5.4.937