Which of the commonly used marker enzymes gives the best results in colorimetric and fluorimetric enzyme immunoassays: Horseradish peroxidase, alkaline phosphatase or β-galactosidase?

Porstmann, B; Porstmann, T; Nugel, E; Evers, U

doi:10.1016/0022-1759(85)90388-6

Cited by 90 publications

(42 citation statements)

References 12 publications

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“…The introduction of specific, high affinity MAbs exhibiting little or no non-specific binding should help to produce more sensitive and reproducible tests. Antibody specificity and to some extent affinity are probably the most significant factors affecting the sensitivity of a sandwich immunoassay (Porstmann et al, 1985;Siddle, 1985). MAbs are especially useful in labelled-antibody (or immunometric) assays because the labelled antibody is present in excess and it is essential to have little or no non-specific binding for maximum sensitivity (Siddle, 1985).…”

Section: Potato Virus Testingmentioning

confidence: 99%

Developments in methodology of plant virus detection

Torrance¹

1992

Netherlands Journal of Plant Pathology

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As a general rule some form of polyclonal-antibody based enzyme immunoassay is still preferred for routine plant virus detection, but modifications may be necessary when increased sensitivity or specificity is required. In recent years new developments in antibody-based detection methods have mostly involved the provision of specific reagents, such as monoclonal antibodies or affinity-purified second antibody-enzyme conjugates which have helped to improve the sensitivity and specificity of the assays. Other kinds of tests (such as nucleic acid hybridisation) must be used when tests for virus coat protein are not appropriate. Recently, tests based on the polymerase chain reaction show great promise. There is no single universal test: assays must be devised and optimised for each pathogen. The challenge with all types of test is to make them quicker, less labour intensive, andif possible -cheaper.

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Section: Potato Virus Testingmentioning

confidence: 99%

Developments in methodology of plant virus detection

Torrance¹

1992

Netherlands Journal of Plant Pathology

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“…The anti-IgG molecules or Protein A were covalently attached to this SAM surface via the bifunctional reagent glutaraldehyde (GA). The aldehyde groups of the GA bind to the amino groups of the protein and form Schiff bases [18]. The immobilization strategy was employed to prepare functional layer with both randomly and specifically oriented antibodies at the sensor surface.…”

Section: Immobilization Proceduresmentioning

confidence: 99%

QCM Immunosensor Based on Polyamidoamine Dendrimers

et al. 2006

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The poly(amidoamine)(PAMAM) dendrimers of fourth generation (G4) and 1-hexadecanethiol (HDT) has been used for formation of self assembled monolayers on a gold surface for preparation of quartz crystal microbalance (QCM) immunosensor for detection of human IgG. The anti-IgG antibodies were immobilized on a surface of G4-HDT layers or those covered by Protein A. In later case the oriented immobilization of anti-IgG takes place due to preferred interaction of F c portion of antibodies with the Protein A. The detection limit in a flow mode was 7 nM both for biosensor with and without Protein A. However, in the case of Protein A the number of bounded Ab was 1.4 times lower in comparison with those without Protein A. Thus, the sensitivity of the Protein A based sensor was higher due to orientation effect in comparison with randomly oriented Ab at a dendrimer surface without Protein A. Increased concentration of IgG and formation of Ab-Ag complexes resulted in increase of motional resistance, that suggest increased contribution of viscoelasticity, as revealed from impedance analysis. The most important property of the immunosensor was its ability for regeneration using 0.1 M glycine-HCl.

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“…That limit is greatly dependent on antibody affinities, in particular, the capture antibody. In general, the signal is amplified with horseradish peroxidase, alkaline phosphatase or β-galactosidase marker enzymes that are conjugated to the detection antibodies [4]. The amplification performed by these nonlinear enzymes improves the sensitivity of the ELISA assay; however, the capability of analyzing low abundant antigens using these enzymes is limited due to "substrate inhibition" [5].…”

Section: Introductionmentioning

confidence: 99%

Non-invasive, ultra-sensitive, high-throughput assays to quantify rare biomarkers in the blood

Freudenberg

Bembas

Greene

et al. 2008

Methods

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Many diseases are easier to treat and control when detected at an early stage of disease progression. Often, disease-related antigens or biomarkers are shed from the primary site and present in the blood. Unfortunately, there are very few tests capable of detecting these rare biomarkers in the blood. A blood test would be very useful to diagnose the disease earlier, monitor effectiveness of treatments, predict recurrence, and monitor recurrence. There is certainly a need to develop assays that are ultrasensitive, non-invasive and high-throughput. Here we describe several highly sensitive immunological assays we have developed to detect rare serum antigens. Initially we created an assay named immunodetection amplified by T7 RNA polymerase (IDAT). To enhance the effectiveness and streamline the procedure, this assay was amended to the facile amplification system termed fluorescent amplification catalyzed by T7 polymerase technique (FACTT). These assays have been used to analyze the tumor antigen HER2 and the prion protein PrP Sc . They can also be applied to other tumor markers or antigens from a variety of diseases such as cardiovascular disease, rheumatoid arthritis, Alzheimer's disease, Parkinson's disease, and hepatitis. These tests are not limited to testing only serum, but may also be applicable to detecting biomarkers in tissue, saliva, urine, cerebrospinal fluid, etc. Clearly, the FACTT-based technology represents an important step in the detection of rare molecules in fluids or tissues for a variety of diseases.

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Which of the commonly used marker enzymes gives the best results in colorimetric and fluorimetric enzyme immunoassays: Horseradish peroxidase, alkaline phosphatase or β-galactosidase?

Cited by 90 publications

References 12 publications

Developments in methodology of plant virus detection

Developments in methodology of plant virus detection

QCM Immunosensor Based on Polyamidoamine Dendrimers

Non-invasive, ultra-sensitive, high-throughput assays to quantify rare biomarkers in the blood

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