Analytical and Bioanalytical Chemistry

, Volume 393, Issue 2, pp 615–621 | Cite as

Frontal analysis microchip capillary electrophoresis to study the binding of ligands to receptors derivatized on magnetic beads

Original Paper


The model binding of the glycopeptide antibiotic teicoplanin (Teic) from Actinoplanes teichomyceticus, immobilized on magnetic microspheres, to d-Ala-d-Ala terminus peptides was assessed using microchip capillary electrophoresis (MCE) with continuous frontal analysis (FA). Teic is closely related to vancomycin (Van), historically, the drug of last resort used to treat many Gram-positive bacterial infections. Glycopeptide antibiotics inhibit bacterial growth by binding to the d-Ala-d-Ala terminus on the cell wall of Gram-positive bacteria via hydrogen bonds, thereby preventing the enzyme-mediated cross-linking of peptidoglycan and eventual cell death. In this work direct and competitive bead-based assays in a microfluidic chip are demonstrated. The binding constants obtained using the technique are comparable with values reported in the literature.


Frontal analysis Magnetic microbeads Glycopeptide antibiotic 


  1. 1.
    Long ZC, Shen Z, Wu DP, Qin JH, Lin BC (2007) Lab Chip 7:1819–1824CrossRefGoogle Scholar
  2. 2.
    Hsiung SK, Lin CH, Lee GB (2005) Electrophoresis 26:1122–1129CrossRefGoogle Scholar
  3. 3.
    Liu CN, Toriello NM, Mathies RA (2006) Anal Chem 78:5474–5479CrossRefGoogle Scholar
  4. 4.
    Lagally ET, Medintz I, Mathies RA (2001) Anal Chem 73:565–570CrossRefGoogle Scholar
  5. 5.
    Liu XJ, Liu X, Liang AY, Shen Z (2006) Electrophoresis 27:3125–3128CrossRefGoogle Scholar
  6. 6.
    Stettler AR, Schwarz MA (2005) J Chromatogr A 1063:217–225CrossRefGoogle Scholar
  7. 7.
    Le Saux T, Hisamoto H, Terabe S (2006) J Chromatogr A 1104:352–358CrossRefGoogle Scholar
  8. 8.
    Chiem NH, Harrison DJ (1998) Electrophoresis 19:3040–3044CrossRefGoogle Scholar
  9. 9.
    Lin YH, Lee GB, Li CW, Huang GR (2001) J Chromatogr A 937:115–125CrossRefGoogle Scholar
  10. 10.
    Chuang YJ, Huang JW, Makamba H, Tsai ML, Li CW, Chen SH (2006) Electrophoresis 27:4158–4165CrossRefGoogle Scholar
  11. 11.
    Bromberg A, Mathies RA (2003) Anal Chem 75:1188–1195CrossRefGoogle Scholar
  12. 12.
    Vlckova M, Stettler AR, Schwarz MA (2006) J Liq Chromatogr Related Technol 29:1047–1076CrossRefGoogle Scholar
  13. 13.
    Gong MJ, Wehmeyer KR, Limbach PA, Heineman WR (2007) Electrophoresis 28:837–842CrossRefGoogle Scholar
  14. 14.
    Liu XJ, Liang AY, Shen Z, Liu X, Zhang Y, Dai ZP, Xiong BH, Lin BC (2006) Electrophoresis 27:5128–5131CrossRefGoogle Scholar
  15. 15.
    Dynabeads@ M-270 Immunoassays (Carboxyl) Manuals, InvitrogenGoogle Scholar
  16. 16.
    Georgieva J, Milling A, Orfanos CE, Geilen CC (2002) Melanoma Res 12:309–317CrossRefGoogle Scholar
  17. 17.
    Gu HW, Ho PL, Tsang KWT, Wang L, Xu B (2003) J Am Chem Soc 125:15702–15703CrossRefGoogle Scholar
  18. 18.
    Fan ZH, Mangru S, Granzow R, Heaney P, Ho W, Dong Q, Kumar R (1999) Anal Chem 71:4851–4859CrossRefGoogle Scholar
  19. 19.
    Jordan A, Scholz R, Maier-Hauff K, van Landeghem F, Waldoefner N, Teichgraeber U, Pinkernelle J, Bruhn H, Neumann F, Thiesen BV, Deimling A, Felix R (2006) J Neurooncol 78:7–14CrossRefGoogle Scholar
  20. 20.
    Hayes MA, Polson NA, Phayre AN, Garcia AA (2001) Anal Chem 73:5896–5902CrossRefGoogle Scholar
  21. 21.
    Duffy DC, McDonald JC, Schueller OJA, Whitesides GM (1998) Anal Chem 70:4974–4984CrossRefGoogle Scholar
  22. 22.
    Wu DP, Zhao BX, Dai ZP, Qin JH, Lin BC (2006) Lab Chip 6:942–947CrossRefGoogle Scholar
  23. 23.
    Kahne D, Leimkuhler C, Wei L, Walsh C (2005) Chem Rev 105:425–448CrossRefGoogle Scholar
  24. 24.
    Loll PJ, Axelsen PH (2000) Annu Rev Biophys, Biomol Struct 29:265–289CrossRefGoogle Scholar
  25. 25.
    Bardone MR, Paternoster M, Coronelli C (1978) J Antibiot 31:276–283Google Scholar
  26. 26.
    Barry AL, Thornsberry C, Jones RN (1986) J Clin Microbiol 23:100–103Google Scholar
  27. 27.
    Chaudhury MKW, Whitesides GM (1991) Langmuir 7:1013–1025CrossRefGoogle Scholar
  28. 28.
    Duffy DC, Mcmonald JC, Schueller OJA, Whitesides GM (1998) Anal Chem 70:4974–4984CrossRefGoogle Scholar
  29. 29.
    Hillborg HA, Gedde UW, Smith GD, Yasuda HK, Wikstrom K (2000) Polymer 41:6851–6863CrossRefGoogle Scholar
  30. 30.
    Huang X, Gordon MJ, Zare RN (1988) Anal Chem 60:1837–1838CrossRefGoogle Scholar
  31. 31.
    Chattejee AN, Perkins HR (1966) Biochem Biophys Res Commun 24:489–494CrossRefGoogle Scholar
  32. 32.
    Perkins HR (1969) Biochem J 111:195–205Google Scholar
  33. 33.
    Nieto M, Perkins HR (1971) Biochem J 123:773–781Google Scholar
  34. 34.
    Piyasena ME, Rea LJ, Diamond RA, Xu H, Gomez FA (2008) Anal Bioanal Chem 392:877–886CrossRefGoogle Scholar
  35. 35.
    Silverio CF, Plazas A, Moran J, Gomez FA (2002) J Liq Chromatogr Related Technol 25:1677–1691CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryCalifornia State University, Los AngelesLos AngelesUSA

Personalised recommendations