Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method.

  • Authors:
    • M Suzuki
    • Y Katsube
    • H Suzuki
    • H Takemura
    • T Sumida
  • View Affiliations

  • Published online on: April 1, 1999     https://doi.org/10.3892/ijmm.3.4.385
  • Pages: 385-475
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Abstract

To examine the relationship between pathophysiological effects and molecular features of anti-double-stranded (ds)DNA autoantibodies (Abs), we isolated anti-dsDNA Ab fragments by using the phage-display method. Fd ç and light-chain DNA were PCR amplified from peripheral blood lymphocytes of a patient with systemic lupus erythematosus (SLE), complicated with lupus nephritis. They were then inserted into a phagemid vector, pComb3-H. We generated eight Fab fragments that specifically bound to solid-phase DNA. The Fab clones stained positively using Crithidia luciliae, indicating that they were anti-dsDNA antibodies. Nucleotide sequences of VH of Fab clones were very similar and appeared to be derived from VH26 germline gene. Differences in the activities of anti-dsDNA Ab of the Fab clones may be ascribed to the diversity of VL. Fab fragments with anti-dsDNA Ab activities exhibited a positive charge on isoelectric focusing, consistent with pathogenic features. Our results indicate that anti-dsDNA Ab Fab fragments obtained by the phage-display method in the present study may possess molecular and functional characteristics of pathogenic anti-dsDNA autoAbs in SLE.

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Apr 1999
Volume 3 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Suzuki M, Katsube Y, Suzuki H, Takemura H and Sumida T: Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method.. Int J Mol Med 3: 385-475, 1999
APA
Suzuki, M., Katsube, Y., Suzuki, H., Takemura, H., & Sumida, T. (1999). Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method.. International Journal of Molecular Medicine, 3, 385-475. https://doi.org/10.3892/ijmm.3.4.385
MLA
Suzuki, M., Katsube, Y., Suzuki, H., Takemura, H., Sumida, T."Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method.". International Journal of Molecular Medicine 3.4 (1999): 385-475.
Chicago
Suzuki, M., Katsube, Y., Suzuki, H., Takemura, H., Sumida, T."Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method.". International Journal of Molecular Medicine 3, no. 4 (1999): 385-475. https://doi.org/10.3892/ijmm.3.4.385