Article Detail

Home > Article Detail
  • P-ISSN 1225-0163
  • E-ISSN 2288-8985

Bioluminescence immunoassay for neurotransmitter, serotonin using aequorin as a Label

Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2010, v.23 no.1, pp.60-67
https://doi.org/10.5806/AST.2010.23.1.060




  • Downloaded
  • Viewed

Abstract

A sensitive competitive heterogeneous bioluminescence immunoassay for serotonin was developed using photoprotein, aequorin as a label for the first time with the optimal assay conditions; especially, serotoninavidin conjugate was prepared by Mannich reaction and the synthetic process of serotonin-avidin conjugate was optimized by controlling the initial molar ratios of serotonin, formaldehyde and avidin (1:12,000:25). The developed bioluminescence immunoassay for serotonin showed good sensitivity (LOD of 0.68 ng/mL) with wide area of dynamic range (5.0×10−10 M~5.0×10−7 M). (cf. the range for serotonin in human blood serum is 151±45 ng/mL). In addition, cross-reactivity studies demonstrated that 5-methoxytryptamine showed some cross-reactivity (28.0%), whereas 3-methylindole, melatonin and 5-hydroxylindole-3-acetic acid showed no crossreactivity,and good recoveries were obtained in serum. Thus, this developed method provides a good tool to monitor serotonin in serum.

keywords
Bioluminescence immunoassay, Aequorin, Neurotransmitter, Serotonin


Reference

1

1. R. F. Masseyeff, W. H. Albert and N. A. Staines, “Methods of Immunological Analysis” Vols. 1 and 2, N. A. Stanines, Ed., Wiley-VCH , NY, U.S.A., 1993.

2

2. E. P. Diamandis and T. K. Christopoulos, “Immunoassay”, Academic Press, San Diego, U.S.A., 1996.

3

3. J. M. Kendall and M. N. Badminton, Trends Biotechnol., 16, 216-224(1998).

4

4. Y. N. Shim and I. R. Paeng, Anal. Sci., 17, a41~a44 (2001).

5

5. S. Shrestha, I. R. Paeng, S. K. Deo and S. Daunert, Bioconjugate Chem., 13, 269-275(2002).

6

6. Y. N. Shim and I. R. Paeng, Bull. Kor. Chem. Soc., 24, 70-74(2003).

7

7. C. H. George, J. M. Kendall, A. K. Campbell and W. H. Evans, J. Biol. Chem., 273, 29822-29829(1998).

8

8. S. Ramanathan, J. C. Lewis, M. S. Kindy and S. Daunert, Anal. Chim. Acta., 369, 181-188(1998).

9

9. C. L. Crofcheck, A. L. Grosvenor, K. W. Anderson, J. K. Lumpp, D. L. Scott and S. Daunert, Anal. Chem., 69, 4768-4772(1997).

10

10. B. Galvan and T. K. Christopoulos, Anal. Chem., 68, 3545-3550(1996).

11

11. D. Prasher, R. O. McCann and M. J. Cormier, Biochem. Biophys. Res. Commun., 126, 1259-1268(1985).

12

12. Y. Kumarasamy, M. Middleton, R. G. Reid, L. Nahar and S. D. Sarker, Fitoterapia, 74, 609-712(2003).

13

13. I. Semak, E. Korik, M. Naumova, J. Wortsman and A. Slominski, Arch. Biochem. Biophysics, 421, 61-66(2004).

14

14. M. Israel, Neurochem. International, 42, 215-220(2003).

15

15. J. Kim, M. Jeon, K.-J. Paeng and I. R. Paeng, Anal. Chim. Acta., 619, 87-93(2008).

16

16. M. Jeon and I. R. Paeng, Ana. Chim. Acta., 619, 180- 185(2008).

17

17. M. Jeon, J. Kim, K.-J. Paeng, S.-W. Park and I. R. Paeng, Microchem. J., 88, 26-31(2008).

18

18. J. Kim, H. Park, J. Ryu, O. Jeon and I. R. Paeng, J. Immunoassay and Immunochemistry 31, 33-44(2010).

19

19. R. Ekins and P. Edwards, Clin. Chem., 43, 1824-1837 (1997).

20

20. Z. D. Peterson, M. L. Lee and S. W. Graves, J. Chromatogr. B., 810, 101-110(2004).

21

21. K. Wu, J. Fei and S. Hu, Anal. Biochem., 318, 100- 106(2003).

상단으로 이동

Analytical Science and Technology