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Development of automated amperometric detection of antibodies against Bacillus anthracis protective antigen

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Abstract

Picogram levels of antibodies against the protective antigen (PA) of Bacillus anthracis were detected in an automated electrochemical sandwich-type enzyme-linked immunosorbant assay. The antibodies were captured and detected using an 8 × 3 array of 50-μm-diameter cavities. The reagent and sample volumes were as low as 200 nL in a less than 25-min assay from capture to signal generation. The electrochemical detection of the antibodies was demonstrated at 0.05–10 μg/mL containing only 10–5,000 pg antibodies. The limit of detection is 10 fg for a 200-nL sample. Detection of anti-PA immunoglobulin G performed in spiked normal human serum and fresh whole human blood did not show a significant difference from detection in a buffer. The initial automation of the assay involved the use of a digital syringe pump for the delivery of reagents to the capture surface.

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References

  1. CDC (2001) Morb Mortal Wkly Rep 50:889–897

    Google Scholar 

  2. Guidi-Rontani C, Mock M (2002) In: Koehler TM (ed) Anthrax. Springer, Berlin, pp 115–141

    Google Scholar 

  3. Inglesby TV, O’Toole T, Henderson DA, Bartlett JG, Ascher MS, Eitzen E, Friedlander AM, Gerberding J, Hauer J, Hughes J, McDade J, Osterholm MT, Parker G, Perl TM, Russell PK, Tonat K (2002) JAMA 287:2236–2252

    Article  Google Scholar 

  4. Walt DR, Franz DR (2000) Anal Chem 72:738A–746A

    Article  CAS  Google Scholar 

  5. Ligler FS, Anderson GP, Davidson PT, Foch RJ, Ives JT, King OK, Page G, Stenger DA, Whelan JP (1998) Environ Sci Technol 32:2461–2466

    Article  CAS  Google Scholar 

  6. Rowe C, Leonard T, Feldstein M, Golden PJ, Scruggs S, MacCraith BD, Cras J, Ligler F (1999) Anal Chem 71:3846–3852

    Article  CAS  Google Scholar 

  7. Rowe-Taitt CA, Golden PJ, Feldstein M, Cras J, Hoffman KE, Ligler F (2000) Biosens Bioelectron 14:785–794

    Article  CAS  Google Scholar 

  8. Rowe-Taitt CA, Haazzard JW, Hoffman KE, Cras J, Golden PJ, Ligler F (2000) Biosens Bioelectron 15:579–589

    Article  CAS  Google Scholar 

  9. McBride MT, Gammon S, Pitesky M, O’Brien TW, Smith T, Aldrich J, Langlois RG, Colston B, Venkateswaran KS (2003) Anal Chem 75:1924–1930

    Article  CAS  Google Scholar 

  10. McBride MT, Masquelier D, Hindson BJ, Makarewicz AJ, Brown S, Burris K, Metz K, Langlois RG, Tsang KW, Bryan R, Anderson DA, Venkateswaran KS, Milanovich FP, Colston BW (2003) Anal Chem 75:5293–5299

    Article  CAS  Google Scholar 

  11. Biagini RE, Sammons DL, Smith JP, MacKenzie BA, Striley CAF, Roberts SA, Snawder JE, Quinn CP (2005) Anal Bioanal Chem 382:1027–1034

    Article  CAS  Google Scholar 

  12. Biagini RE, Sammons DL, Smith JP, MacKenzie BA, Striley CAF, Semenova S, Steward-Clark E, Stamey K, Freeman AE, Quinn CP, Snawder JE (2004) Clin Diagn Lab Immunol 11:50–55

    Article  CAS  Google Scholar 

  13. Quinn CP, Semenova VA, Elie CM, Romero-Steiner S, Greene C, Li H, Stamey K, Steward-Clark E, Schmidt DS, Mothershed E, Pruckler J, Schwartz S, Benson RF, Helsel LO, Holder PF, Johnson SE, Kellum M, Messmer T, Thacker WL, Besser L, Plikaytis LD, Taylor TH Jr, Freeman E, Wallace KJ, Dull P, Sejvar J, Bruce E, Moreno R, Schuchat A, Lingappa JR, Marano N, Martin SK, Walls J, Bronsdon M, Carlone GM, Bajani-Ari M, Ashford DA, Stephens DS, Perkins BA (2002) Emerg Infect Dis 8:1103–1110

    CAS  Google Scholar 

  14. Aguilar ZP, Vandaveer WR, Fritsch I (2002) Anal Chem 74:3321–3329

    Article  CAS  Google Scholar 

  15. Henry C, Fritsch F (1999) Anal Chem 71:550–556

    Article  CAS  Google Scholar 

  16. Henry C, Fritsch I (1999) J lectrochem Soc 9:146:3367–3373

    Google Scholar 

  17. Aguilar ZP, Van Nguyen C, Sirisena M, Gertsch J, Arumugam P, Spencer D, Wansapura C, Aguilar Y, Homesley J (2006) ECS Trans 3(10):125–137

    Article  CAS  Google Scholar 

  18. Everett R, Fritsch-Faules I (1995) Anal Chim Acta 307:253–268

    Article  CAS  Google Scholar 

  19. Everett WR, Welch TL, Reed L, Fritsch-Faules I (1995) Anal Chem 67:292–298

    Article  CAS  Google Scholar 

  20. Schoenfish MH, Pemberton JE (1998) J Am Chem Soc 120:4502–4513

    Article  Google Scholar 

  21. Norrod KL, Rowlen KL (1998) J Am Chem Soc 120:2656–2657

    Article  CAS  Google Scholar 

  22. Butler JE (1991) Immunochemistry of solid phase immunoassay, 1st edn. CRC, Boca Raton, p 319

    Google Scholar 

  23. Jiang L, Glidle A, Griffith A, McNeil CJ, Cooper JM (1997) Bioelectrochem Bioenerg 42:15–23

    Article  CAS  Google Scholar 

  24. Maggio ET (1980) Enzyme immunoassay. CRC, Boca Raton

    Google Scholar 

  25. Zull JE, Reed-Mundell J, Lee YW, Vezenov D, Ziats NP, Anderson JM, Sukenik CN (1994) J Ind Microbiol 13:137–143

    Article  CAS  Google Scholar 

  26. Tang HT, Lunte CE, Halsall HB, Heineman WR (1988) Anal Chim Acta 214:187–195

    Article  CAS  Google Scholar 

  27. Xu Y, Halsall HB, Heineman WR (1989) J Pharm Biomed Anal 7:1301–1311

    Article  CAS  Google Scholar 

  28. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

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Acknowledgements

We are grateful to the National Science Foundation for financial support (NSF DMI no. 0441594). We thank the University of Arkansas for use of equipment and facilities in the Department of Chemistry and Biochemistry and for use of the microfabrication facilities at the High Density Electronics Center. We also thank the University of Arkansas Statewide Mass Spectrometry Facility for analysis of PAPP. We acknowledge Emily Clark and Prabhu Arumugam for fabrication of the chips, Jana Gertsch for maintenance of laboratory supplies, and Dmitri Brevnov for discussions.

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  1. Vegrandis, LLC, 700 W. Research Ctr. Blvd., ENRC 1208, Fayetteville, AR, 72701, USA

    Zoraida P. Aguilar & Mali Sirisena

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  1. Zoraida P. Aguilar
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  2. Mali Sirisena
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Correspondence to Zoraida P. Aguilar.

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Aguilar, Z.P., Sirisena, M. Development of automated amperometric detection of antibodies against Bacillus anthracis protective antigen. Anal Bioanal Chem 389, 507–515 (2007). https://doi.org/10.1007/s00216-007-1448-z

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  • DOI: https://doi.org/10.1007/s00216-007-1448-z

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