Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 2637 - 2642 |
| Number of pages | 6 |
| Journal | Analytical Chemistry |
| Volume | 77 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2005 |
| MoE publication type | A1 Journal article-refereed |
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Keywords
- immunoassay
- antibodies
- detection
- drug detection
- drug abuse
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One-step homogeneous immunoassay for small analytes. / Pulli, Timo (Corresponding Author); Höyhtyä, Matti; Söderlund, Hans; Takkinen, Kristiina.
In: Analytical Chemistry, Vol. 77, No. 8, 2005, p. 2637 - 2642.Research output: Contribution to journal › Article › Scientific › peer-review
TY - JOUR
T1 - One-step homogeneous immunoassay for small analytes
AU - Pulli, Timo
AU - Höyhtyä, Matti
AU - Söderlund, Hans
AU - Takkinen, Kristiina
PY - 2005
Y1 - 2005
N2 - We have developed a one-step, homogeneous noncompetitive immunoassay for small analytes using recombinant antibodies and morphine as the model analyte. A highly specific antibody against the immune complex (IC) formed between an anti-morphine antibody and morphine was selected from a naïve scFv phage display library. The in vitro phage library selection procedure avoids the difficulties associated with the production of anti-IC antibodies by animal immunization. The anti-morphine and the anti-IC antibodies were labeled with a pair of fluorescence resonance energy transfer (FRET) fluorophores. In the FRET assay the labeled antibodies were incubated with saliva samples spiked with morphine, codeine, or heroin. Within 2 min, 5 ng/mL morphine, which is clearly under the recommended cutoff level, was detected without cross-reactivity to codeine or heroin. This assay principle is also widely applicable to other small analytes.
AB - We have developed a one-step, homogeneous noncompetitive immunoassay for small analytes using recombinant antibodies and morphine as the model analyte. A highly specific antibody against the immune complex (IC) formed between an anti-morphine antibody and morphine was selected from a naïve scFv phage display library. The in vitro phage library selection procedure avoids the difficulties associated with the production of anti-IC antibodies by animal immunization. The anti-morphine and the anti-IC antibodies were labeled with a pair of fluorescence resonance energy transfer (FRET) fluorophores. In the FRET assay the labeled antibodies were incubated with saliva samples spiked with morphine, codeine, or heroin. Within 2 min, 5 ng/mL morphine, which is clearly under the recommended cutoff level, was detected without cross-reactivity to codeine or heroin. This assay principle is also widely applicable to other small analytes.
KW - immunoassay
KW - antibodies
KW - detection
KW - drug detection
KW - drug abuse
U2 - 10.1021/ac048379l
DO - 10.1021/ac048379l
M3 - Article
VL - 77
SP - 2637
EP - 2642
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 8
ER -