Abstract
A novel protocol for the synthesis of dye-encapsulating liposomes tagged with DNA oligonucleotides at their outer surface was developed. These liposomes were optimized for use as signal enhancement agents in lateral-flow sandwich-hybridization assays for the detection of single-stranded RNA and DNA sequences. Liposomes were synthesized using the reverse-phase evaporation method and tagged with oligonucleotides by adding cholesteryl-modified DNA probes to the initial lipid mixture. This resulted in a greatly simplified protocol that provided excellent control of the probe coverage on the liposomes and cut the preparation time from 16 hours to just 6 hours. Liposomes were prepared using probe concentrations ranging from 0.00077 to 0.152 mol% of the total lipid, several hydrophobic and polyethylene glycol-based spacers between the cholesteryl anchor and the probe, and liposome diameters ranging from 208 nm to 365 nm. The liposomes were characterized by dynamic light scattering, visible spectroscopy, and fluorescence spectroscopy. Their signal enhancement functionality was compared by using them in lateral-flow optical biosensors for the detection of single-stranded DNA sequences. In these assays, an optimal reporter probe concentration of 0.013 mol%, liposome diameter of 315 nm, and liposome optical density of 0.4–0.6 at 532 nm were found. The spacer length between the cholesteryl anchor and the probe showed no significant effect on the signals in the lateral-flow assays. The results presented here provide important data for the general use of liposomes as labels in analytical assays, with specific emphasis on nucleic acid detection via lateral flow assays.
Similar content being viewed by others
Abbreviations
- Cholesteryl-TEG:
-
1-dimethoxytrityl-3-O-(N-cholesteryl-3-aminopropyl)-triethyleneglycolyl-glyceryl-2-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite
- DPPC:
-
1,2-dipalmitoyl-sn-glycero-3-phosphocholine
- DPPG:
-
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)]
- NASBA:
-
nucleic acid sequence based amplification
- SATA:
-
N-succinimidyl S-acetylthioacetate
- SRB:
-
sulforhodamine B
- Sulfo-KMUS:
-
N-[κ-maleimidoundecanoyloxy]sulfosuccinimide ester
References
Zhou X, Huang L, Li S (2001) Biosens Bioelectron 16:85–95
Iqbal S, Mayo M, Bruno J, Bronk B, Batt C, Chambers J (2001) Biosens Bioelectron 15:549–578
Kricka L (2002) Ann Clin Biochem 39:114–129
Kricka L (1999) Clin Chem 45:453–458
Rule G, Montagna R, Durst R (1997) Anal Biochem 244:260–269
Rule G, Montagna R, Durst R (1996) Clin Chem 42:1206–1209
Baeumner A, Cohen R, Miksic V, Min J (2003) Biosens Bioelectron 18:405–413
Baeumner A, Schlesinger N, Slutzki N, Romano J, Lee EM, Montagna RA (2002) Anal Chem 74:1442–1448
Hartley H (2002) A rapid and sensitive Bacillus anthracis biosensor. MS Thesis, Cornell University, Ithaca, NY
Baeumner A, Pretz J, Fang S (2004) Anal Chem 76:888–894
Robinson A, Creeth J, Jones M (1998) Biochim Biophys Acta 1369:278–286
Jones M, Kilpatrick P, Carbonell R (1993) Biotechnol Prog 9:242–258
Jones M, Kilpatrick P, Carbonell R (1994) Biotechnol Prog 10:174–186
Plant A, Brizgys M, Locascio-Brown L, Durst R (1989) Anal Biochem 176:420–426
Rongen H, van der Horst H, Hugenholtz G, Bult A, Bennekom W, van der Meide P (1994) Anal Chim Acta 287:191–199
Rongen H, van Nierop T, van der Horst H, Rombouts R, van der Meide P, Bult A, Bennekom W (1995) Anal Chim Acta 306:333–341
Esch M, Locascio L, Tarlov M, Durst R (2001) Anal Chem 73:2952–2958
Zaytseva N, Montagna R, Lee E, Baeumner A (2005) Anal Bioanal Chem 380:46–53
Ho J, Durst R (2000) Anal Chim Acta 414:51–60
Chen C, Baeumner A, Durst R (2005) Talanta 67:205–211
Barbet J, Machet P, Leserman L (1981) J Supramol Struct Cell Biochem 16:243–258
Martin FJ, Heath TD (1992) Covalent attachment of proteins to liposomes. In: New RRC (ed) Liposomes, a practical approach. IRL Press/Oxford University Press, Oxford, pp 163–182
Zhang N, Ping Q, Huang G, Xu W (2005) Int J Pharm 294:247–259
Kung V, Redemann C (1986) Biochem Biophys Acta 862:435–439
Hansen C, Yao G, Moase E, Zalipsky S, Allen T (1995) Biochim Biophys Acta 1239:133–144
Hartley H, Baeumner A (2003) Anal Bioanal Chem 376:319–327
Wen H, DeCory T, Borejsza-Wysocki W, Durst R (2006) Talanta 68:1264–1272
Bredehorst R, Ligler F, Kusterbeck A, Chang E, Gaber B, Vogel C (1986) Biochemistry 25:5693–5698
Xu C, Taylor P, Ersoz M, Fletcher P, Paunov V (2003) J Mater Chem 13:3044–3048
Henderson GB, Stein CA (1995) Nucleic Acids Res 23:3726–3731
Letsinger RL, Zhang G, Sun DK, Ikeuchi T, Sarin PS (1989) Proc Natl Acad Sci USA 86:6553–6556
Krieg A, Tonkinson J, Matson S, Zhao Q, Saxon M, Zhang L, Bhanja U, Yakubov L, Stein C (1993) Proc Natl Acad Sci USA 90:1048–1052
Bartlett G (1959) J Biol Chem 234:466–468
Small D (1967) J Lipid Res 8:551–557
Israelachvili J, Mitchell D (1975) Biochim Biophys Acta 389:13–19
Fiske C, SubbaRow Y (1925) J Biol Chem 66:375–399
Esch M, Baeumner A, Durst R (2001) Anal Chem 73:3162–3167
Limanto J, Tallarico J, Porter J, Khuong K, Houk K, Snapper M (2002) J Am Chem Soc 124:14748–14758
Muller G, Schmidt B, Jiřiček J, Bünzli J, Schenk K (2003) Acta Crystal C 59:o353–o356
Berman H, Olson W, Beveridge D, Westbrook J, Gelbin A, Demeny T, Hsieh S, Srinivasan A, Schneider B (1992) Biophys J 63:751–759
Mrevlishvili G, Kankia B, Mdzinarashvili T, Brelidze T, Khvedelidze M, Metreveli N, Razmadze G (1998) Chem Phys Lipids 94:139–143
Huang W, Zhang Z, Han X, Tang J, Wang J, Dong S, Wang E (2003) Bioelectrochemistry 59:21–27
Acknowledgements
The authors thank Dr. John Randolph of Glen Research Corporation for calculating the lengths of the spacers used in this study. We also thank Dr. John C. March for critically reviewing this manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Edwards, K.A., Baeumner, A.J. Optimization of DNA-tagged dye-encapsulating liposomes for lateral-flow assays based on sandwich hybridization. Anal Bioanal Chem 386, 1335–1343 (2006). https://doi.org/10.1007/s00216-006-0705-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-006-0705-x