Abstract
We report the analysis of human rhinovirus serotype 2 (HRV2) on a commercially available lab‐on‐a‐chip instrument. Due to lack of sufficient native fluorescence, the proteinaceous capsid of HRV2 was labeled with Cy5 for detection by the red laser (λ ex 630 nm) implemented in the instrument. On the microdevice, electrophoresis of the labeled virus was possible in a BGE without stabilizing detergents, which is in contrast to conventional CE; moreover, analysis times were drastically shortened to the few 10 s range. Resolution of the sample constituents (virions, a contaminant present in all virus preparations, and excess dye) was improved upon adaptation of the separation conditions, mainly by adjusting the SDS concentration of the BGE. Purity of fractions from size‐exclusion chromatography after labeling of virus was assessed, and affinity complex formation of the labeled virus with various recombinant very‐low‐density lipoprotein receptor derivatives differing in the number of concatenated V3 ligand binding repeats was monitored. Virus analysis on microchip devices is of particular interest for experiments with infectious material because of easy containment and disposal of samples. Thus, the employment of microchip devices in routine analysis of viruses appears to be exceptionally attractive.
Keywords: Chip electrophoresis, Concatemer, Cy5, HRV2, Lab‐on‐a‐chip, Virus
References
- 1. Liu, D. , Shi, M. , Huang, H. , Long, Z. et al., J. Chromatogr. B 2006, 844, 32–38. [DOI] [PubMed] [Google Scholar]
- 2. Chen, Y.‐H. , Wang, W.‐C. , Young, K.‐C. , Chang, T.‐Y. , Chen, S.‐H. , Clin. Chem. 1999, 45, 1938–1943. [PubMed] [Google Scholar]
- 3. Huang, F.‐C. , Liao, C.‐S. , Lee, G.‐B. , Electrophoresis 2006, 27, 3297–3305. [DOI] [PubMed] [Google Scholar]
- 4. Zhou, X. , Liu, D. , Zhong, R. , Dai, Z. et al., Electrophoresis 2004, 25, 3032–3039. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5. Sanders, J. C. , Breadmore, M. C. , Mitchell, P. S. , Landers, J. P. , Analyst 2002, 127, 1558–1563. [DOI] [PubMed] [Google Scholar]
- 6. Hofgartner, W. T. , Huhmer, A. F. R. , Landers, J. P. , Kant, J. A. , Clin. Chem. 1999, 45, 2120–2128. [PubMed] [Google Scholar]
- 7. Grom, F. , Kentsch, J. , Mueller, T. , Schnelle, T. , Stelzle, M. , Electrophoresis 2006, 27, 1386–1393. [DOI] [PubMed] [Google Scholar]
- 8. Vegvari, A. , Hjertén, S. , Electrophoresis 2003, 24, 3815–3820. [DOI] [PubMed] [Google Scholar]
- 9. Okun, V. M. , Ronacher, B. , Blaas, D. , Kenndler, E. , Anal. Chem. 1999, 71, 2028–2032. [DOI] [PubMed] [Google Scholar]
- 10. Okun, V. M. , Blaas, D. , Kenndler, E. , Anal. Chem. 1999, 71, 4480–4485. [DOI] [PubMed] [Google Scholar]
- 11. Okun, V. , Ronacher, B. , Blaas, D. , Kenndler, E. , Anal. Chem. 2000, 72, 2553–2558. [DOI] [PubMed] [Google Scholar]
- 12. Okun, V. M. , Ronacher, B. , Blaas, D. , Kenndler, E. , Anal. Chem. 2000, 72, 4634–4639. [DOI] [PubMed] [Google Scholar]
- 13. Okun, V. M. , Moser, R. , Ronacher, B. , Kenndler, E. , Blaas, D. , J. Biol. Chem. 2001, 276, 1057–1062. [DOI] [PubMed] [Google Scholar]
- 14. Okun, V. M. , Moser, R. , Blaas, D. , Kenndler, E. , Anal. Chem. 2001, 73, 3900–3906. [DOI] [PubMed] [Google Scholar]
- 15. Okun, V. M. , Nizet, S. , Blaas, D. , Kenndler, E. , Electrophoresis 2002, 23, 896–902. [DOI] [PubMed] [Google Scholar]
- 16. Kremser, L. , Okun, V. M. , Nicodemou, A. , Blaas, D. , Kenndler, E. , Anal. Chem. 2004, 76, 882–887. [DOI] [PubMed] [Google Scholar]
- 17. Kremser, L. , Konecsni, T. , Blaas, D. , Kenndler, E. , Anal. Chem. 2004, 76, 4175–4181. [DOI] [PubMed] [Google Scholar]
- 18. Kremser, L. , Petsch, M. , Blaas, D. , Kenndler, E. , Anal. Chem. 2004, 76, 7360–7365. [DOI] [PubMed] [Google Scholar]
- 19. Nicodemou, A. , Petsch, M. , Konecsni, T. , Kremser, L. et al., FEBS Lett. 2005, 579, 5507–5511. [DOI] [PubMed] [Google Scholar]
- 20. Bilek, G. , Kremser, L. , Blaas, D. , Kenndler, E. , Electrophoresis 2006, 27, 3999–4007. [DOI] [PubMed] [Google Scholar]
- 21. Kremser, L. , Petsch, M. , Blaas, D. , Kenndler, E. , Electrophoresis 2006, 27, 1112–1121. [DOI] [PubMed] [Google Scholar]
- 22. Kremser, L. , Petsch, M. , Blaas, D. , Kenndler, E. , Electrophoresis 2006, 27, 2630–2637. [DOI] [PubMed] [Google Scholar]
- 23. Bilek, G. , Kremser, L. , Wruss, J. , Blaas, D. , Kenndler, E. , Anal. Chem. 2007, 79, 1620–1625. [DOI] [PubMed] [Google Scholar]
- 24. Hewat, E. A. , Neumann, E. , Conway, J. F. , Moser, R. et al., EMBO J. 2000, 19, 6317–6325. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25. Ronacher, B. , Marlovits, T. C. , Moser, R. , Blaas, D. , Virology 2000, 278, 541–550. [DOI] [PubMed] [Google Scholar]
- 26. Moser, R. , Snyers, L. , Wruss, J. , Angulo, J. et al., Virology 2005, 338, 259–269. [DOI] [PubMed] [Google Scholar]
- 27. Weiss, V. U. , Kolivoska, V. , Kremser, L. , Gaš, B. et al., in press.
- 28. Bousse, L. , Mouradian, S. , Minalla, A. , Yee, H. et al., Anal. Chem. 2001, 73, 1207–1212. [DOI] [PubMed] [Google Scholar]
- 29. Verdaguer, N. , Blaas, D. , Fita, I. , J. Mol. Biol. 2000, 300, 1179–1194. [DOI] [PubMed] [Google Scholar]
- 30. Neumann, E. , Moser, R. , Snyers, L. , Blaas, D. , Hewat, E. A. , J. Virol. 2003, 77, 8504–8511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31. Verdaguer, N. , Fita, I. , Reithmayer, M. , Moser, R. , Blaas, D. , Nat. Struct. Mol. Biol. 2004, 11, 429–434. [DOI] [PubMed] [Google Scholar]
- 32. Konecsni, T. , Kremser, L. , Snyers, L. , Rankl, C. et al., FEBS Lett. 2004, 568, 99–104. [DOI] [PubMed] [Google Scholar]
- 33. Vlasak, M. , Roivainen, M. , Reithmayer, M. , Goesler, I. et al., J. Virol. 2005, 79, 7389–7395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 34. Wruss, J. , Ruenzler, D. , Steiger, C. , Chiba, P. et al., Biochemistry 2007, 46, 6331–6339. [DOI] [PubMed] [Google Scholar]