Chromatographic Immunoassays

Hage, David S.; Moser, Annette C.

doi:10.1201/9780824751982.ch29

Cited by 17 publications

(66 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many of these detection methods require the use of an antibody or target analog that contains a chemical label or enzymatic tag. However, some of these methods make use of the inherent properties of the target for its direct detection [7,10]. This latter situation often occurs in the case of absorbance detection.…”

Section: Detection Methods In Chromatographic Immunoassaysmentioning

confidence: 99%

“…Common examples are absorbance and fluorescence detection, with chemiluminescence, electrochemical detection, radiometric detection, thermal measurements and MS also having been employed [7,8,10]. Many of these detection methods require the use of an antibody or target analog that contains a chemical label or enzymatic tag.…”

Section: Detection Methods In Chromatographic Immunoassaysmentioning

confidence: 99%

“…Antibodies are glycoproteins that are produced by the immune system in response to a foreign agent, or antigen [2,3]. Antibodies can be generated against a wide range of compounds and can have both specific and strong binding, with many antibody-target interactions having association equilibrium constants in the range of 10 5 to 10 12 M -1…”

mentioning

confidence: 99%

“…Various binding agents, detection methods, supports and assay formats have been reported for this group of methods. In addition, this set of techniques has been used in applications ranging from low mass compounds such as drugs, hormones and herbicides to higher mass targets such as peptides, proteins and bacteria [6][7][8][9][10][11][12][13][14].…”

mentioning

confidence: 99%

“…One method that has been of interest in recent years is a chromatographic immunoassay or immunochromatographic assay (also sometimes called a 'flow immunoassay' or 'flowinjection immunoassay') [5][6][7][8][9][10][11][12][13]. In this technique, an antibody or antibody-related agent (e.g., an antibody fragment) is attached to a chromatographic support or used as part of a chromatographic system for the isolation or measurement of a specific target.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

et al. 2015

View full text Add to dashboard Cite

show abstract

Section: Detection Methods In Chromatographic Immunoassaysmentioning

confidence: 99%

Section: Detection Methods In Chromatographic Immunoassaysmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

et al. 2015

View full text Add to dashboard Cite

show abstract

Chromatographic competitive binding immunoassays: a comparison of the sequential and simultaneous injection methods

Nelson

Reiter

Hage

2003

Biomedical Chromatography

View full text Add to dashboard Cite

Two approaches for performing competitive binding immunoassays by HPLC and other flow-based systems are the simultaneous and sequential injection methods. Both these techniques make use of a column with a limited amount of antibody, onto which is injected a sample and a fixed amount of a labeled analyte analog. An indirect measure of the unlabeled analyte in the sample is then obtained by looking at the amount of analog in either the nonretained or retained peaks. In the simultaneous injection mode, the sample and labeled analog are applied at the same time to the column, while in the sequential mode the sample is injected first, followed by the analog. This results in a difference in the analytical characteristics of these two approaches. This study used chromatographic theory and previous data obtained for injections of human serum albumin (HSA) onto an anti-HSA antibody column to compare the response, detection limits, range, and sensitivity of these methods. Under equivalent conditions, it was found that the sequential method always provided the best lower limit of detection and sensitivity. However, the simultaneous mode had a broader dynamic range and higher upper limit of detection. From these observations, several guidelines were developed regarding the use and selection of such assays for new applications.

show abstract

Immunoaffinity Chromatography for Protein Purification and Analysis

Sharmeen,

Suh,

Kyei

et al. 2023

Current Protocols

View full text Add to dashboard Cite

Immunoaffinity chromatography (IAC) is a type of liquid chromatography that uses immobilized antibodies or related binding agents as selective stationary phases for sample separation or analysis. The strong binding and high selectivity of antibodies have made IAC a popular tool for the purification and analysis of many chemicals and biochemicals, including proteins. The basic principles of IAC are described as related to the use of this method for protein purification and analysis. The main factors to consider in this technique are also presented under a discussion of the general strategy to follow during the development of a new IAC method. Protocols, as illustrated using human serum albumin (HSA) as a model protein, are provided for the use of IAC in several formats. This includes both the use of IAC with traditional low‐performance supports such as agarose for off‐line immunoextraction and supports used in high‐performance IAC for on‐line immunoextraction. The use of IAC for protein analysis as a flow‐based or chromatographic immunoassay is also discussed and described using HSA and a competitive binding assay format as an example. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Off‐line immunoextraction by traditional immunoaffinity chromatographyBasic Protocol 2: On‐line immunoextraction by high‐performance immunoaffinity chromatographyBasic Protocol 3: Competitive binding chromatographic immunoassay

show abstract

Chromatographic Immunoassays

Cited by 17 publications

References 0 publications

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

Chromatographic competitive binding immunoassays: a comparison of the sequential and simultaneous injection methods

Immunoaffinity Chromatography for Protein Purification and Analysis

Contact Info

Product

Resources

About