A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction

  • Authors:
    • Xiaoqin Fan
    • Jinmei Wei
    • Haiting Xiong
    • Xiaohui Liu
    • Serge Benichou
    • Xuejuan Gao
    • Langxia Liu
  • View Affiliations

  • Published online on: August 25, 2015     https://doi.org/10.3892/ijo.2015.3132
  • Pages: 1485-1493
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The human immunodeficiency virus (HIV) protein negative factor (Nef) is important for AIDS pathogenesis. An anti-Nef single-domain antibody (sdAb19) derived from camelids has been previously generated and shown to effectively block the physiological functions of Nef in vitro and in vivo in nef-transgenic mice. However, sdAb19 must be ectopically expressed within the target cell to be able to exert its neutralizing effect on Nef, while the extra-cellular administration method turned out to be ineffective. This might suggest a default of the stability or/and deliverability of sdAb19. The identification of small molecule compounds capable of inhibiting the Nef-sdAb19 interaction and mimicking the neutralizing activity of sdAb19 in vivo would therefore be the means of circumventing the problem encountered with sdAb19. Here we describe the development of a high-throughput screening method combining the homogeneous time-resolved fluorescence (HTRF) and the microscale thermophoresis (MST) techniques for the identification of small-molecule compounds inhibiting the Nef-sdAb19 interaction by binding to Nef protein. Eight small-molecule compounds have been selected for their ability to significantly inhibit the Nef-sdAb19 interaction and to bind to Nef. These molecules could be further assessed for their potential of being the Nef-neutralizing agents in the future.
View Figures
View References

Related Articles

Journal Cover

October-2015
Volume 47 Issue 4

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

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Fan X, Wei J, Xiong H, Liu X, Benichou S, Gao X and Liu L: A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. Int J Oncol 47: 1485-1493, 2015
APA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., & Liu, L. (2015). A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. International Journal of Oncology, 47, 1485-1493. https://doi.org/10.3892/ijo.2015.3132
MLA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47.4 (2015): 1485-1493.
Chicago
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47, no. 4 (2015): 1485-1493. https://doi.org/10.3892/ijo.2015.3132