376:
356:
402:, free analyte analog molecules labeled with an enzyme (e.g., glucose-6-phosphate dehydrogenase enzyme) compete with the analyte being tested. The active enzyme reduces NAD (no signal) to NADH (which absorbs at 340 nm), so absorbance is monitored at 340 nm. When the labeled analyte binds to the Ab, the enzyme becomes inactive, and a signal is generated by the free label. The signal intensity is directly proportional to the analyte concentration.
245:
40:
421:, free antibodies bind to drug microparticle conjugates to form aggregates that absorb in the visible range in the absence of the analyte. In the presence of the analyte, the Ab binds to the free analyte, preventing microparticle aggregation and causing a reduction in absorbance. The signal is inversely proportional to the analyte concentration.
217:
264:. These enzymes allow for detection often because they produce an observable color change in the presence of certain reagents. In some cases these enzymes are exposed to reagents which cause them to produce light or chemiluminescence. There are several types of ELISA: direct, indirect, sandwich, competitive.
127:
The use of a calibrator is often employed in immunoassays. Calibrators are solutions that are known to contain the analyte in question, and the concentration of that analyte is generally known. Comparison of an assay's response to a real sample against the assay's response produced by the calibrators
391:
measures the fluorescence polarization signal after incubation, without separating bound and free labels. Free labeled analyte analog molecules are added to the sample, and their
Brownian motion differs when bound to a large antibody (Ab) versus free in solution. The analyte competes for binding to
467:
The analyte in the unknown sample is bound to the antibody site, then the labelled antibody is bound to the analyte. The amount of labelled antibody on the site is then measured. It will be directly proportional to the concentration of the analyte because the labelled antibody will not bind if the
431:
involves genetically engineering an enzyme (e.g., beta-galactosidase) into two inactive fragments: a small enzyme donor (ED) conjugated with the drug analog, and a larger enzyme acceptor (EA). When the two fragments associate, the full enzyme converts a substrate into a cleaved colored product. If
152:
In addition to the binding of an antibody to its antigen, the other key feature of all immunoassays is a means to produce a measurable signal in response to the binding. Most, though not all, immunoassays involve chemically linking antibodies or antigens with some kind of detectable label. A large
123:
Immunoassays come in many different formats and variations. Immunoassays may be run in multiple steps with reagents being added and washed away or separated at different points in the assay. Multi-step assays are often called separation immunoassays or heterogeneous immunoassays. Some immunoassays
371:
In a competitive, homogeneous immunoassay, unlabelled analyte in a sample competes with labeled analyte to bind an antibody. The amount of labelled, unbound analyte is then measured. In theory, the more analyte in the sample, the more labelled analyte gets displaced and then measured; hence, the
409:
generates singlet oxygen species in microbeads coupled to the analyte, and when the analyte binds to the respective Ab molecule, coupled to another kind of bead, the analyte reacts with singlet oxygen, generating chemiluminescence signals proportional to the concentration of the analyte-Ab
437:
458:
Mixing a sample with labelled antibodies, the targeted analyte is bound by labelled antibodies. The unbound, labelled antibodies are washed away, and the bound, labelled antibodies are measured. The intensity of the signal is directly proportional to the amount of analyte in the sample.
449:
As in a competitive, homogeneous immunoassay, unlabelled analyte in a sample competes with labelled analyte to bind an antibody. In the heterogeneous assays, the labelled, unbound analyte is separated or washed away, and the remaining labelled, bound analyte is measured.
560:
tags. Illuminated by a modulated light at a plasmon resonance wavelength, the nanoparticles generate strong acoustic signal, which can be measured using a microphone. The photoacoustic immunoassay can be applied to lateral flow tests, which use colloidal nanoparticles.
1567:
Tsivou M; Kioukia-Fougia N; Lyris E; Aggelis Y; Fragkaki A; Kiousi X; Simitsek Ph; Dimopoulou H; Leontiou I-P; Stamou M; Spyridaki M-H; Georgakopoulos C (2006). "An overview of the doping control analysis during the
Olympic Games of 2004 in Athens, Greece".
153:
number of labels exist in modern immunoassays, and they allow for detection through different means. Many labels are detectable because they either emit radiation, produce a color change in a solution, fluoresce under light, or can be induced to emit light.
338:
While some kind of label is generally employed in immunoassays, there are certain kinds of assays which do not rely on labels, but instead employ detection methods that do not require the modification or labeling the components of the assay.
432:
drug analyte molecules are present, they compete with the ED-labeled drug in solution for the limited Ab sites. Free ED-labeled drug analog will bind to EA, generating a colorimetric signal directly proportional to the amount of analyte.
136:
Immunoassays rely on the ability of an antibody to recognize and bind a specific macromolecule in what might be a complex mixture of macromolecules. In immunology the particular macromolecule bound by an antibody is referred to as an
148:
In some cases, an immunoassay may use an antigen to detect for the presence of antibodies, which recognize that antigen, in a solution. In other words, in some immunoassays, the analyte may be an antibody rather than an antigen.
112:, although it may be other kinds of molecules, of different sizes and types, as long as the proper antibodies that have the required properties for the assay are developed. Analytes in biological liquids such as
1099:
Zanut, A.; Fiorani, A.; Canola, S.; Saito, T.; Ziebart, N.; Rapino, S.; Rebeccani, S.; Barbon, A.; Irie, T.; Josel, H.; Negri, F.; Marcaccio, M.; Windfuhr, M.; Imai, K.; Valenti, G.; Paolucci, F. (2020).
198:. This type of immunoassay is now used in around 100 million clinical tests every year worldwide, enabling clinicians to measure a wide range of proteins, pathogens and other molecules in blood samples.
343:
is an example of technique that can detect binding between an unlabeled antibody and antigens. Another demonstrated labeless immunoassay involves measuring the change in resistance on an electrode as
124:
can be carried out simply by mixing the reagents and samples and making a physical measurement. Such assays are called homogeneous immunoassays, or less frequently non-separation immunoassays.
1705:
213:
Immunoassays employ a variety of different labels to allow for detection of antibodies and antigens. Labels are typically chemically linked or conjugated to the desired antibody or antigen.
885:
283:
RIAs were some of the earliest immunoassays developed, but have fallen out of favor largely due to the difficulty and potential dangers presented by working with radioactivity.
175:
Immunoassays became considerably simpler to perform and more popular when techniques for chemically linked enzymes to antibodies were demonstrated in the late 1960s.
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468:
analyte is not present in the unknown sample. This type of immunoassay is also known as a sandwich assay as the analyte is "sandwiched" between two antibodies.
1512:
611:
363:
Immunoassays can be run in a number of different formats. Generally, an immunoassay will fall into one of several categories depending on how it is run.
1691:
392:
the Ab, and if the labeled analyte binds to the Ab, a signal is produced. The signal intensity is inversely proportional to the analyte concentration.
1291:
814:
Aydin, Suleyman (2015). "A short history, principles, and types of ELISA, and our laboratory experience with peptide/protein analyses using ELISA".
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525:
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238:
375:
917:
44:
Illustration of the basic components of an immunoassay, which includes an analyte (green), an antibody (black), and a detectable label (yellow)
1391:
440:
Two-site, noncompetitive immunoassays usually consist of an analyte "sandwiched" between two antibodies. ELISAs are often run in this format.
386:
1630:
Zhao Y, Huang Y, Zhao X, McClelland JF, Lu M (2016). "Nanoparticle-based photoacoustic analysis for highly sensitive lateral flow assays".
889:
742:
292:
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295:(RT qPCR) and traditional immunoassay techniques. Called real-time immunoquantitative PCR (iqPCR) the label used in these assays is a
684:
359:
In a competitive, homogeneous immunoassay unlabeled analyte displaces bound labelled analyte, which is then detected or measured.
426:
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Georgios
Tsekenis (2008). "Label-free immunosensor assay for myelin basic protein based upon an ac impedance protocol".
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makes it possible to interpret the signal strength in terms of the presence or concentration of analyte in the sample.
1894:
498:
1200:
J. B. González-Díaz; et al. (2008). "Plasmonic Au/Co/Au nanosandwiches with
Enhanced Magneto-Optical Activity".
970:
Gofflot; El (2004). "Immuno-quantitative polymerase chain reaction for detection and quantitation of prion protein".
1102:"Insights into the mechanism of coreactant electrochemiluminescence facilitating enhanced bioanalytical performance"
280:(RIA). Radioactivity emitted by bound antibody-antigen complexes can be easily detected using conventional methods.
1942:
913:"Immunoquantitative real-time PCR for detection and quantification of Staphylococcus aureus enterotoxin B in foods"
799:
1527:
179:
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524:, and some are also used for the detection and/or quantitative measurement of some pharmaceutical compounds (see
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50:
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Immunoassays are used in sports anti-doping laboratories to test athletes' blood samples for prohibited
355:
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Other clinical immunoassays are quantitative; they measure amounts. Immunoassays can measure levels of
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17,000,000,000 and was thought to have prospects of slow annual growth in the 2 to 3 percent range.
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1853:
1731:
1419:"Detection of early pregnancy forms of human chorionic gonadotropin by home pregnancy test devices"
1395:
1595:
Zhao Y, Cao M, McClelland JF, Lu M (2016). "A photoacoustic immunoassay for biomarker detection".
1819:
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768:
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183:
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for her work in immunoassays in 1977, becoming the second
American woman to have won the award.
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Homogeneous competitive assays: FPIA, EMIT, LOCI, KIMS and CEDIA. See section text for details.
1906:
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are credited with the development of the first immunoassays in the 1950s. Yalow accepted the
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493:. More specifically, they are qualitative tests that detect whether hCG is present, using a
372:
amount of labelled, unbound analyte is proportional to the amount of analyte in the sample.
277:
233:. Immunoassays which employ enzymes are referred to as enzyme immunoassays (EIAs), of which
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The photoacoustic immunoassay measures low-frequency acoustic signals generated by metal
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1460:
Guglielmi G (September 2020). "Fast coronavirus tests: what they can and can't do".
999:
843:
1874:
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688:
641:
Yetisen A. K. (2013). "Paper-based microfluidic point-of-care diagnostic devices".
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307:
113:
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330:(ECL), in which the label emits detectable light in response to electric current.
104:(sometimes). The molecule detected by the immunoassay is often referred to as an "
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85:
17:
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1342:"On the Slow Diffusion of Point-of-Care Systems in Therapeutic Drug Monitoring"
1125:
1911:
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are frequently measured using immunoassays for medical and research purposes.
1358:
1680:
Chapter 5 and 6 in the book "Bioanalytical
Chemistry" by Susan R. Mikkelsen
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679:. In "Biographical Memoirs". National Academy of Sciences 1990;59:54-71.
97:
1666:"The Immunoassay Handbook", 3rd Edition, David Wild, Ed., Elsevier,2008
39:
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1222:
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are immunoassays, called immunodiagnostics in this context. Many home
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101:
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703:"Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA)"
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and the area on an antigen to which the antibody binds is called an
318:
are a type of immunoassay that often employ fluorogenic reporters.
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581:
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243:
234:
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Possibly one of the most popular labels to use in immunoassays is
215:
117:
27:
Biochemical test for a protein or other molecule using an antibody
1040:"Protein microarray on-demand: a novel protein microarray system"
531:
Drug testing also starts with a quick qualtitative immunoassay.
419:
particle enhanced turbidimetric inhibition immunoassay (PETINIA)
216:
1687:
296:
1293:
Fluorescence
Applications in Biotechnology and Life Sciences
276:
can be incorporated into immunoassay reagents to produce a
859:"ELISA test marks 35 years of answering medical questions"
96:
or a small molecule in a solution through the use of an
415:
kinetic interaction of microparticle in solution (KIMS)
489:
are immunoassays, which detect the pregnancy marker
291:
A newer approach to immunoassays involves combining
201:
By 2012, the commercial immunoassay industry earned
1862:
1828:
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49:
32:
407:luminescent oxygen channeling immunoassay (LOCI)
293:real-time quantitative polymerase chain reaction
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1333:
1331:
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1327:
1325:
1699:
888:. America.gov. April 27, 2008. Archived from
314:are used in a number of modern immunoassays.
8:
803:. Vol. 34, no. 4. pp. 12–13.
248:ELISA plate showing various cortisol levels
1706:
1692:
1684:
972:Journal of Immunoassay and Immunochemistry
612:Surround Optical Fiber Immunoassay (SOFIA)
501:is also a qualitative, lateral-flow test.
220:A sandwich ELISA run on a microtitre plate
38:
1673:at the U.S. National Library of Medicine
1436:
1367:
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1221:
1133:
1075:
1065:
946:
718:
1552:"Principles of Drug Testing Technology,"
775:, Royal Society of Chemistry (RSC), 2003
374:
1800:Enzyme multiplied immunoassay technique
633:
526:Enzyme multiplied immunoassay technique
445:Competitive, heterogeneous immunoassays
398:enzyme multiplied immunoassay technique
239:enzyme multiplied immunoassay technique
918:Applied and Environmental Microbiology
29:
1513:"Immunoassays, applications: Clinica"
1159:Annual Review of Analytical Chemistry
463:Two-site, noncompetitive immunoassays
454:One-site, noncompetitive immunoassays
387:fluorescence polarization immunoassay
367:Competitive, homogeneous immunoassays
7:
1179:10.1146/annurev-anchem-060908-155305
795:"Seizing Immunoassay Opportunities"
793:Carlson, Bruce (15 February 2014).
88:test that measures the presence or
62:
1750:Ouchterlony double immunodiffusion
1520:Encyclopedia of Analytical Science
241:(EMIT) are the most common types.
235:enzyme-linked immunosorbent assays
25:
857:Susan J. Landers (3 April 2006).
1557:, Retrieved online Dec 07, 2012.
541:recombinant human growth hormone
1392:"ELISA for Home Pregnancy Test"
427:cloned enzyme donor immunoassay
252:Enzymes used in ELISAs include
1038:Chatterjee; Sitaraman (2008).
828:10.1016/j.peptides.2015.04.012
744:Prof Anthony Campbell - MA PhD
1:
1740:Chromatin immunoprecipitation
1597:Biosensors and Bioelectronics
911:Rajkovic; El-Moualij (2006).
1785:Chemiluminescent immunoassay
1765:Counterimmunoelectrophoresis
1067:10.1371/journal.pone.0003265
720:10.1373/clinchem.2005.051532
491:human chorionic gonadotropin
322:Electrochemiluminescent tags
190:used in immunoassay with an
1895:Direct fluorescent antibody
1438:10.1093/clinchem/47.12.2131
535:Sports anti-doping analysis
499:COVID-19 rapid antigen test
351:Classifications and formats
1959:
1609:10.1016/j.bios.2016.05.028
1482:10.1038/d41586-020-02661-2
1340:Sanavio B, Krol S (2015).
1290:Goldys, Ewa (2009-08-24).
1126:10.1038/s41467-020-16476-2
800:Gen. Eng. Biotechnol. News
194:that makes its own light:
1917:Total complement activity
1582:10.1016/j.aca.2005.08.068
1526:: 317–324. Archived from
1014:"Luminex xMAP Technology"
552:Photoacoustic Immunoassay
518:prostate-specific antigen
508:to assess heart disease,
341:Surface plasmon resonance
108:" and is in many cases a
63:
37:
1880:Complement fixation test
1675:Medical Subject Headings
1359:10.3389/fbioe.2015.00020
617:CD/DVD based immunoassay
328:electrochemiluminescence
1346:Front Bioeng Biotechnol
886:"Rosalyn Sussman Yalow"
334:Label-free immunoassays
1755:Radial immunodiffusion
1570:Analytica Chimica Acta
1214:10.1002/smll.200700594
543:(rhGH, rGH, hGH, GH).
441:
380:
360:
254:horseradish peroxidase
249:
221:
1870:Diagnostic immunology
1760:Immunoelectrophoresis
1018:Millipore Corporation
984:10.1081/ias-200028044
864:American Medical News
439:
378:
358:
326:Some labels work via
303:Fluorogenic reporters
247:
219:
162:Rosalyn Sussman Yalow
1890:Immunohistochemistry
1398:on 21 September 2017
1319:License: "CC BY 4.0"
1256:Analytical Chemistry
939:10.1128/AEM.03068-05
602:Magnetic immunoassay
268:Radioactive isotopes
258:alkaline phosphatase
1885:Immunocytochemistry
1854:Latex fixation test
1732:Immunoprecipitation
1638:(46): 19204–19210.
1511:Twyman, SA (2001).
1474:2020Natur.585..496G
1417:Butler, SA (2001).
1171:2009ARAC....2..359F
1118:2020NatCo..11.2668Z
1058:2008PLoSO...3.3265C
931:2006ApEnM..72.6593R
892:on October 20, 2012
528:for more details).
316:Protein microarrays
178:In 1983, Professor
1820:Immunofluorescence
1815:Radiobinding assay
1644:10.1039/C6NR05312B
1424:Clinical Chemistry
749:Cardiff University
655:10.1039/C3LC50169H
442:
381:
361:
250:
222:
188:radioactive iodine
184:Cardiff University
1943:Immunologic tests
1930:
1929:
1907:Skin allergy test
1555:Drug Tests Direct
1468:(7826): 496–498.
1269:10.1021/ac702070e
701:Lequin R (2005).
677:Solomon A. Berson
649:(12): 2210–2251.
622:Agglutination-PCR
597:Lateral flow test
196:chemiluminescence
74:
73:
16:(Redirected from
1950:
1838:Hemagglutination
1810:Radioimmunoassay
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1533:on 24 March 2012
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607:Radioimmunoassay
481:A wide range of
278:radioimmunoassay
192:acridinium ester
180:Anthony Campbell
100:(usually) or an
67:edit on Wikidata
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18:Immunoreactivity
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1300:. p. 311.
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422:
411:
403:
393:
368:
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304:
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288:
285:
269:
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226:
223:
210:
207:
166:Solomon Berson
158:
155:
133:
130:
72:
71:
64:
61:
60:
53:
47:
46:
43:
35:
34:
26:
24:
14:
13:
10:
9:
6:
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3:
2:
1955:
1944:
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1896:
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1873:
1871:
1868:
1867:
1865:
1861:
1855:
1852:
1848:
1845:
1844:
1843:
1842:Hemagglutinin
1839:
1836:
1835:
1833:
1831:
1830:Agglutination
1827:
1821:
1818:
1816:
1813:
1811:
1808:
1806:
1803:
1801:
1798:
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1715:Medical tests
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1019:
1015:
1009:
1006:
1001:
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993:
989:
985:
981:
978:(3): 241–58.
977:
973:
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954:
949:
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932:
928:
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920:
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904:
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796:
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764:
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708:
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697:
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690:
686:
685:0-309-04198-8
682:
678:
672:
669:
664:
660:
656:
652:
648:
644:
643:Lab on a Chip
637:
634:
628:
623:
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483:medical tests
476:
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366:
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333:
331:
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321:
319:
317:
313:
312:phycoerythrin
309:
302:
300:
298:
294:
287:DNA reporters
286:
284:
281:
279:
275:
267:
265:
263:
259:
255:
246:
242:
240:
237:(ELISAs) and
236:
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176:
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131:
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125:
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107:
103:
99:
95:
94:macromolecule
91:
90:concentration
87:
83:
79:
68:
58:
54:
52:
48:
41:
36:
31:
19:
1875:Nephelometry
1776:
1725:86000–86849)
1665:
1635:
1631:
1625:
1600:
1596:
1590:
1573:
1569:
1562:
1554:
1547:
1535:. Retrieved
1528:the original
1523:
1519:
1506:
1465:
1461:
1455:
1428:
1422:
1412:
1400:. Retrieved
1396:the original
1386:
1349:
1345:
1311:. Retrieved
1292:
1285:
1260:
1254:
1248:
1208:(2): 202–5.
1205:
1201:
1195:
1162:
1158:
1152:
1109:
1105:
1094:
1052:(9): e3265.
1049:
1043:
1033:
1021:. Retrieved
1008:
975:
971:
965:
922:
916:
906:
894:. Retrieved
890:the original
880:
868:. Retrieved
862:
852:
819:
815:
809:
798:
788:
777:, retrieved
772:
763:
752:, retrieved
743:
737:
710:
706:
696:
676:
671:
646:
642:
636:
592:Nephelometry
558:nanoparticle
555:
538:
530:
514:hypoglycemia
503:
495:lateral flow
480:
466:
457:
448:
425:
418:
414:
406:
396:
385:
370:
362:
347:bind to it.
337:
325:
306:
290:
282:
272:Radioactive
271:
251:
228:
212:
200:
177:
174:
160:
151:
147:
135:
126:
122:
81:
77:
75:
1847:Coombs test
1777:Immunoassay
1671:Immunoassay
1537:11 December
1402:11 December
1313:11 December
1223:10261/17402
1112:(1): 2668.
1106:Nat. Commun
1023:13 December
779:29 December
769:"NPS Focus"
754:29 December
497:setup. The
308:Fluorogenic
170:Nobel Prize
86:biochemical
78:immunoassay
33:Immunoassay
1912:Patch test
1719:immunology
1603:: 261–66.
1165:: 359–85.
870:9 December
707:Clin. Chem
629:References
520:to detect
512:to assess
1805:RAST test
1632:Nanoscale
1498:221768935
1240:206490102
675:Rall JE.
186:replaced
132:Principle
1937:Category
1717:used in
1652:27834971
1617:27183276
1576:: 1–13.
1490:32939084
1447:11719477
1378:25767794
1277:18260654
1232:18196506
1187:20636067
1144:32472057
1086:18813342
1045:PLOS ONE
1000:37548553
992:15461386
957:17021210
896:June 26,
844:36486495
836:25908411
822:: 4–15.
816:Peptides
729:16179424
689:Fulltext
663:23652632
565:See also
547:Research
472:Examples
410:complex.
345:antigens
274:isotopes
260:(AP) or
98:antibody
1795:ELISpot
1470:Bibcode
1369:4341557
1167:Bibcode
1135:7260178
1114:Bibcode
1077:2533396
1054:Bibcode
948:1610299
927:Bibcode
510:insulin
429:(CEDIA)
413:In the
395:In the
299:probe.
256:(HRP),
231:enzymes
225:Enzymes
157:History
143:epitope
139:antigen
110:protein
106:analyte
102:antigen
84:) is a
57:D007118
1922:MELISA
1677:(MeSH)
1650:
1615:
1496:
1488:
1462:Nature
1445:
1376:
1366:
1352:: 20.
1304:
1275:
1238:
1230:
1185:
1142:
1132:
1084:
1074:
998:
990:
955:
945:
842:
834:
727:
683:
661:
577:MELISA
400:(EMIT)
389:(FPIA)
209:Labels
1863:Other
1790:ELISA
1531:(PDF)
1516:(PDF)
1494:S2CID
1236:S2CID
1202:Small
996:S2CID
840:S2CID
582:ECLIA
572:ELISA
506:CK-MB
118:urine
114:serum
92:of a
65:[
1648:PMID
1613:PMID
1539:2012
1486:PMID
1443:PMID
1404:2012
1374:PMID
1315:2012
1302:ISBN
1273:PMID
1228:PMID
1183:PMID
1140:PMID
1082:PMID
1025:2012
988:PMID
953:PMID
898:2010
872:2012
832:PMID
781:2012
756:2012
725:PMID
681:ISBN
659:PMID
424:The
417:and
405:The
384:The
203:US$
164:and
51:MeSH
1723:CPT
1640:doi
1605:doi
1578:doi
1574:555
1478:doi
1466:585
1433:doi
1364:PMC
1354:doi
1265:doi
1218:hdl
1210:doi
1175:doi
1130:PMC
1122:doi
1072:PMC
1062:doi
980:doi
943:PMC
935:doi
824:doi
715:doi
651:doi
297:DNA
182:at
116:or
76:An
1939::
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1634:.
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1601:85
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1429:47
1427:.
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1261:80
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994:.
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951:.
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830:.
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797:.
771:,
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723:.
711:51
709:.
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1212::
1206:4
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1177::
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1056::
1050:3
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1002:.
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959:.
937::
929::
900:.
874:.
846:.
826::
731:.
717::
691:.
665:.
653::
80:(
69:]
20:)
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