Abstract
The practice of immunoassay has experienced a widespread transition from radioisotopic labeling to nonisotopic labeling over the last two decades. Radioisotope labels have drawbacks that hamper their applications: (i) perceived radiation hazards of reagents, (ii) regulatory requirements and disposal problems of working with radioactive materials, and (iii) short shelf-life of the labeled reagents. The advantage of isotopic labeling is the incorporation into analytes without altering structure or reactivity, as is often the case with ELISA or fluorescent detection systems. We developed a format for isotope label immunoassay with the long-life isotope 14C as the label and accelerator mass spectrometer (AMS) as the detection system. AMS quantifies attomole levels of several isotopes, including 14C. With this exquisite sensitivity, the sensitivity of an immunoassay is limited by the K(d) of the antibody and not the detection system. The detection limit of the assays for atrazine and 2,3,7,8-tetrachlorodibenzo-p-dioxin was 2.0 x 10- 10 M and 2.0 x 10-11 M, respectively, approximately an order of magnitude below the standard enzyme immunoassay. Notably, < 1 dpm (0.45 pCi)of 14C-labeled compound was used in each assay, which is well below the limit of disposal (50 nCi per g) as nonradioactive waste. Thus, endogenous reporter ligands quantified by AMS provide the advantages of an RIA without the associated problems of radioactive waste.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2445-2449 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 97 |
| Issue number | 6 |
| DOIs | |
| State | Published - Mar 14 2000 |
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ASJC Scopus subject areas
- Genetics
- General
Cite this
Isotope-labeled immunoassays without radiation waste. / Shan, Guomin; Huang, Wei; Gee, Shirley J.; Buchholz, Bruce A.; Vogel, John S.; Hammock, Bruce D.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 6, 14.03.2000, p. 2445-2449.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Isotope-labeled immunoassays without radiation waste
AU - Shan, Guomin
AU - Huang, Wei
AU - Gee, Shirley J.
AU - Buchholz, Bruce A.
AU - Vogel, John S.
AU - Hammock, Bruce D.
PY - 2000/3/14
Y1 - 2000/3/14
N2 - The practice of immunoassay has experienced a widespread transition from radioisotopic labeling to nonisotopic labeling over the last two decades. Radioisotope labels have drawbacks that hamper their applications: (i) perceived radiation hazards of reagents, (ii) regulatory requirements and disposal problems of working with radioactive materials, and (iii) short shelf-life of the labeled reagents. The advantage of isotopic labeling is the incorporation into analytes without altering structure or reactivity, as is often the case with ELISA or fluorescent detection systems. We developed a format for isotope label immunoassay with the long-life isotope 14C as the label and accelerator mass spectrometer (AMS) as the detection system. AMS quantifies attomole levels of several isotopes, including 14C. With this exquisite sensitivity, the sensitivity of an immunoassay is limited by the K(d) of the antibody and not the detection system. The detection limit of the assays for atrazine and 2,3,7,8-tetrachlorodibenzo-p-dioxin was 2.0 x 10- 10 M and 2.0 x 10-11 M, respectively, approximately an order of magnitude below the standard enzyme immunoassay. Notably, < 1 dpm (0.45 pCi)of 14C-labeled compound was used in each assay, which is well below the limit of disposal (50 nCi per g) as nonradioactive waste. Thus, endogenous reporter ligands quantified by AMS provide the advantages of an RIA without the associated problems of radioactive waste.
AB - The practice of immunoassay has experienced a widespread transition from radioisotopic labeling to nonisotopic labeling over the last two decades. Radioisotope labels have drawbacks that hamper their applications: (i) perceived radiation hazards of reagents, (ii) regulatory requirements and disposal problems of working with radioactive materials, and (iii) short shelf-life of the labeled reagents. The advantage of isotopic labeling is the incorporation into analytes without altering structure or reactivity, as is often the case with ELISA or fluorescent detection systems. We developed a format for isotope label immunoassay with the long-life isotope 14C as the label and accelerator mass spectrometer (AMS) as the detection system. AMS quantifies attomole levels of several isotopes, including 14C. With this exquisite sensitivity, the sensitivity of an immunoassay is limited by the K(d) of the antibody and not the detection system. The detection limit of the assays for atrazine and 2,3,7,8-tetrachlorodibenzo-p-dioxin was 2.0 x 10- 10 M and 2.0 x 10-11 M, respectively, approximately an order of magnitude below the standard enzyme immunoassay. Notably, < 1 dpm (0.45 pCi)of 14C-labeled compound was used in each assay, which is well below the limit of disposal (50 nCi per g) as nonradioactive waste. Thus, endogenous reporter ligands quantified by AMS provide the advantages of an RIA without the associated problems of radioactive waste.
UR - http://www.scopus.com/inward/record.url?scp=0034646463&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034646463&partnerID=8YFLogxK
U2 - 10.1073/pnas.040575997
DO - 10.1073/pnas.040575997
M3 - Article
C2 - 10706612
AN - SCOPUS:0034646463
VL - 97
SP - 2445
EP - 2449
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 6
ER -