Construction of phosphorylatable chimeric monoclonal antibody CC49.

  • Authors:
    • L Lin
    • S D Gillies
    • Y Lan
    • L Izotova
    • W Wu
    • J Schlom
    • S Pestka
  • View Affiliations

  • Published online on: July 1, 1998     https://doi.org/10.3892/ijo.13.1.115
  • Pages: 115-135
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Abstract

Phosphorylation sites were introduced into chimeric monoclonal antibody CC49 (MAb-chCC49) by inserting synthetic fragments encoding two and six phosphorylation sites into an expression vector, pdHL7. The phosphorylation sites were created by using the predicted consensus sequences for phosphorylation by the cAMP-dependent protein kinase to the carboxyl terminus of the heavy chain constant region of the MAb-chCC49. The resultant modified antibodies (MAb-chCC49K1 and MAb-chCC49-6P) were expressed in NS0 cells and purified. The MAb-chCC49K1 protein contains two phosphorylation sites per heavy chain whereas the MAb-chCC49-6P protein contains six sites per heavy chain. Both MAb-chCC49K1 and MAb-chCC49-6P proteins can be phosphorylated by the catalytic subunit of cAMP-dependent protein kinase with [gamma-32P]ATP to high specific activity. The 32P-labeled MAb-chCC49K1 and MAb-chCC49-6P proteins bind to cells expressing TAG-72 antigens. The introduction of phosphorylation sites into a monoclonal antibody provides a reagent for the diagnosis and treatment of cancer. The use of multiple phosphorylation sites provides antibodies with very high specific radioactivity and demonstrates that cassettes of phosphorylation sites can be introduced into proteins without altering their functional activity.

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Jul 1998
Volume 13 Issue 1

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

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Spandidos Publications style
Lin L, Gillies S, Lan Y, Izotova L, Wu W, Schlom J and Pestka S: Construction of phosphorylatable chimeric monoclonal antibody CC49.. Int J Oncol 13: 115-135, 1998.
APA
Lin, L., Gillies, S., Lan, Y., Izotova, L., Wu, W., Schlom, J., & Pestka, S. (1998). Construction of phosphorylatable chimeric monoclonal antibody CC49.. International Journal of Oncology, 13, 115-135. https://doi.org/10.3892/ijo.13.1.115
MLA
Lin, L., Gillies, S., Lan, Y., Izotova, L., Wu, W., Schlom, J., Pestka, S."Construction of phosphorylatable chimeric monoclonal antibody CC49.". International Journal of Oncology 13.1 (1998): 115-135.
Chicago
Lin, L., Gillies, S., Lan, Y., Izotova, L., Wu, W., Schlom, J., Pestka, S."Construction of phosphorylatable chimeric monoclonal antibody CC49.". International Journal of Oncology 13, no. 1 (1998): 115-135. https://doi.org/10.3892/ijo.13.1.115