Clinical chemistry

Stinshoff, K.; Stein, W.; Wood, W. G.; Laska, P. F.

doi:10.1021/ac00139a018

Cited by 5 publications

(5 citation statements)

References 170 publications

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“…The limits of detection for serum and standards were about 1 pmol and 0.2 fmol, respectively (i.e., 10'8-10"n M for a 30-mL sample) (9). Although these results are a clear indication of the potential of HPIAC in automated sandwich immunoassays, the limits of detection reported are comparable to those already obtained with current automated systems (3,4). For the determination of lower concentration analytes, this study will examine the use of HPIAC with an alternative means of detection, namely chemiluminescence.…”

Section: Introductionsupporting

confidence: 79%

“…To overcome these disadvantages in other types of immunoassays, a number of automated systems have been developed. Examples include instruments based on fluorescence polarization, rate nephelometry, particle counting, and EMIT centrifugal analysis (3). These automated techniques, however, have important limitations that prevent their use in sandwich immunoassays.…”

Section: Introductionmentioning

confidence: 99%

“…These automated techniques, however, have important limitations that prevent their use in sandwich immunoassays. For example, all of these systems are based on homogeneous methods, in which simple solution reactions between antibody and analyte are used for compound detection (3,4). As a result, they are not designed to carry out the more complex washing and separation steps required in a sandwich immunoassay.…”

Section: Introductionmentioning

confidence: 99%

“…As a result, they are not designed to carry out the more complex washing and separation steps required in a sandwich immunoassay. Another problem is that these methods typically have limits of detection of 1(T10 M or greater (3,4). In comparison, some compounds determined by sandwich immunoassays may have sample concentrations of less than 10-12 M, requiring much lower detection limits (5).…”

Section: Introductionmentioning

confidence: 99%

“…Like many other trace biological compounds, no system has previously been described for the automated determination of PTH (3). In this work, such a system was developed using an HPIAC column containing immobilized anti-(44-68 PTH) antibodies for extraction of PTH from samples and acridinium ester-labeled anti-(l-34 PTH) antibodies for the hormone's detection.…”

Section: Introductionmentioning

confidence: 99%

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High-performance immunoaffinity chromatography and chemiluminescent detection in the automation of a parathyroid hormone sandwich immunoassay

Hage¹,

Kao²

1991

Anal. Chem.

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An automated sandwich immunoassay was developed based on high-performance immunoaffinity chromatography and chemiluminescent detection, using the determination of parathyroid hormone (PTH) in plasma as a model system. In this method, injections of plasma and acridinium ester-labeled anti-(1-34 PTH) antibodies were made onto a column containing immobilized anti-(44-68 PTH) antibodies. Upon elution, PTH and its associated labeled antibody were combined with an alkaline peroxide postcolumn reagent, and the resulting light production was measured. Factors considered in optimizing this system included the column's dissociation properties, the rate of light production in the postcolumn reactor, and the use of sequential vs simultaneous injection of sample and labeled antibody. The final system developed required 6 min per plasma injection, following a 1-h incubation of sample with labeled antibody. The response was linear over 2-3 orders of magnitude and the lower limit of detection for a 66-microL plasma sample was only 16 amol, or 2.4 x 10(-13) M. Overall, this method had a precision and response similar to those of manual PTH methods but required 24-fold less time to perform. By using different immobilized and labeled antibodies, this method could easily be adapted for use with other analytes.

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Section: Introductionsupporting

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

High-performance immunoaffinity chromatography and chemiluminescent detection in the automation of a parathyroid hormone sandwich immunoassay

Hage¹,

Kao²

1991

Anal. Chem.

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Biopolymers, Analytical Techniques

Ward

2000

Kirk-Othmer Encyclopedia of Chemical Technology

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Analytical techniques that utilize biopolymers, ie, natural macromolecules such as proteins, nucleic acids, and polysaccharides that compose living substances, represent a rapidly expanding field. The number of applications is large, eg, chiral chromatography, immunology, and biosensors, among others. Direct chiral resolution by liquid chromatography (lc) involves diastereomeric interactions between the chiral solute and the chiral stationary phase. Because biopolymers are chiral molecules and can form diastereomeric interactions with chiral solutes, they are ideal for use as chiral stationary phases. This property has led to a rapid growth of chromatographic stationary phases utilizing biopolymers to separate chiral molecules. Biopolymers are employed in many immunological techniques, including the analysis of food, clinical samples, pesticides, and in other areas of analytical chemistry. Immunoassays are specific, sensitive, relatively easy to perform, and usually inexpensive. One condition that must be met for the application of an immunochemical method is the stimulating of an immune response leading to the formation of antibodies in the immunized animal. These antibodies can then be isolated and used as highly specific analytical reagents (immunoassays). Analytes that can combine with the corresponding antibodies are called antigens. In general, large, rigid, chemically complex molecules make good antigens. Antibodies are proteins, found in many body fluids such as tears, saliva, and urine, that are present in highest concentrations in blood serum. The particular proteins which exhibit antibody activity are the immunoglobulins (Ig). Biologically derived polymers can be used as highly selective immobilized reagents in analytical applications. The immobilized enzyme electrode is the most common immobilized biopolymer sensor, consisting of a thin layer of enzyme immobilized on the surface of an electrochemical sensor. The advantages of immobilized enzyme electrodes include minimal pretreatment of the sample matrix, small sample volume, and the recovery of the enzyme for repeated use. Enzyme immunosensors are enzyme immunoassays coupled with electrochemical sensors. Enantiomeric separations are expected to continue to have a considerable economic impact on the development of new drugs and therapy in the biomedical field. The importance of immunoassays for food monitoring and in the detection of diseases is expected to continue to grow. The development of biosensors is expected to benefit monitoring therapeutic drug levels, office testing, and implantable devices because of the advantages of cost‐saving automation and improved data handling.

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Biopolymers

Ward¹

2005

Van Nostrand's Encyclopedia of Chemistry

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Clinical chemistry

Cited by 5 publications

References 170 publications

High-performance immunoaffinity chromatography and chemiluminescent detection in the automation of a parathyroid hormone sandwich immunoassay

High-performance immunoaffinity chromatography and chemiluminescent detection in the automation of a parathyroid hormone sandwich immunoassay

Biopolymers, Analytical Techniques

Biopolymers

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