Screening of Beta-Secretase Inhibitors by Capillary Electrophoresis-Mass Spectrometry

  • Jan SchejbalEmail author
  • Roman Řemínek
  • Zdeněk Glatz
Part of the Methods in Molecular Biology book series (MIMB, volume 2089)


Alzheimer’s disease is the most common cause of dementia, currently afflicting almost 40 million patients worldwide. According to the amyloid cascade hypothesis, the pathogenesis of the disease could be slowed down or even stopped by the inhibition of beta-secretase, making this aspartic acid protease a potentially important drug target site. Capillary electrophoresis is a promising technique for screening putative enzyme inhibitors due to highly effective separations, minuscule sample and other chemicals consumption, compatibility with a variety of detection techniques, and high throughput via automation. This chapter presents a method based on capillary electrophoresis coupled to mass spectrometry detection for kinetic and inhibition assays of the beta-secretase reaction with a decapeptide derived from an amyloid precursor protein.

Key words

Capillary electrophoresis Mass spectrometry Alzheimer’s disease Beta-secretase Enzyme inhibition Screening 



This work was supported by grant no. GA16-06106S from the Czech Science Foundation.


  1. 1.
    Alzheimer’s Association (2015) Alzheimer’s disease facts and figures. Alzheimers Dement 11:332–384CrossRefGoogle Scholar
  2. 2.
    Hardy J, Selkoe DJ (2002) The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297:353–356CrossRefGoogle Scholar
  3. 3.
    Selkoe DJ (2001) Alzheimer’s disease: genes, proteins, and therapy. Physiol Rev 81:741–766CrossRefGoogle Scholar
  4. 4.
    Eikelenboom P, Veerhuis R, Scheper W, Rozemuller AJM, van Gool WA, Hoozemans JJM (2006) The significance of neuroinflammation in understanding Alzheimer’s disease. J Neural Transm 113:1685–1695CrossRefGoogle Scholar
  5. 5.
    Vassar R (2001) The beta-secretase, BACE: a prime drug target for Alzheimer’s disease. J Mol Neurosci 17:157–170CrossRefGoogle Scholar
  6. 6.
    Vassar R, Kuhn PH, Haass C, Kennedy ME, Rajendran L, Wong PC, Lichtenthaler SF (2014) Function, therapeutic potential and cell biology of BACE proteases: current status and future prospects. J Neurochem 130:4–28CrossRefGoogle Scholar
  7. 7.
    Querfurth HW, LaFerla FM (2010) Alzheimer’s disease. N Engl J Med 362:329–344CrossRefGoogle Scholar
  8. 8.
    Glatz Z (2006) Determination of enzymatic activity by capillary electrophoresis. J Chromatogr B 841:23–37CrossRefGoogle Scholar
  9. 9.
    Fan Y, Scriba GK (2010) Advances in-capillary electrophoretic enzyme assays. J Pharm Biomed Anal 53:1076–1090CrossRefGoogle Scholar
  10. 10.
    Scriba GK, Abromeit H, Hense M, Fan Y (2013) Capillary electrophoretic enzyme assays. Methods Mol Biol 984:285–308CrossRefGoogle Scholar
  11. 11.
    Ouimet CM, D’Amico CI, Kennedy RT (2017) Advances in capillary electrophoresis and the implications for drug discovery. Expert Opin Drug Discov 12:213–224CrossRefGoogle Scholar
  12. 12.
    Haass C, Lemere CA, Capell A, Citron M, Seubert P, Schenk D, Lannfelt L, Selkoe DJ (1995) The Swedish mutation causes early-onset Alzheimer’s disease by β-secretase cleavage within the secretory pathway. Nat Med 1:1291–1296CrossRefGoogle Scholar
  13. 13.
    Schejbal J, Slezáčková L, Řemínek R, Glatz Z (2017) A capillary electrophoresis-mass spectrometry based method for the screening of β-secretase inhibitors as potential Alzheimer’s disease therapeutics. J Chromatogr A 1487:235–241CrossRefGoogle Scholar
  14. 14.
    Řemínek R, Slezáčková L, Schejbal J, Glatz Z (2017) Development and comprehensive comparison of two on-line capillary electrophoretic methods for β-secretase inhibitor screening. J Chromatogr A 1518:89–96CrossRefGoogle Scholar
  15. 15.
    Pietrak BL, Crouthamel M-C, Tugusheva K, Lineberger JE, Xu M, DiMuzio JM, Steele T, Espeseth AS, Stachel SJ, Coburn CA, Graham SL, Vacca JP, Shi X-P, Simon AJ, Hazuda DJ, Lai M-T (2005) Biochemical and cell-based assays for characterization of BACE-1 inhibitors. Anal Biochem 342:144–151CrossRefGoogle Scholar
  16. 16.
    Mancini F, De Simone A, Andrisano V (2011) Beta-secretase as a target for Alzheimer’s disease drug discovery: an overview of in vitro methods for characterization of inhibitors. Anal Bioanal Chem 400:1979–1996CrossRefGoogle Scholar
  17. 17.
    May PC, Willis BA, Lowe SL, Dean RA, Monk SA, Cocke PJ, Audia JE, Boggs LN, Borders AR, Brier RA (2015) The potent BACE1 inhibitor LY2886721 elicits robust central Aβ pharmacodynamic responses in mice, dogs, and humans. J Neurosci 35:1199–1210CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2020

Authors and Affiliations

  1. 1.Faculty of Science, Department of BiochemistryMasaryk UniversityBrnoCzech Republic

Personalised recommendations