Theoretical concepts of on-line liquid chromatographic-biochemical detection systems. II. Detection systems based on labelled affinity proteins

Journal of Chromatography. a
A J OostercampJ van der Greef

Abstract

A theoretical concept for on-line liquid chromatography-biochemical detection (LC-BCD) using labelled affinity proteins as reporter molecules is presented. The BCD system is based on the post-column addition of labelled affinity proteins such as fluorescein-labelled streptavidin to the LC effluent. After a short reaction time, free and analyte-bound label are separated during passage through a column packed with an immobilised-ligand support. The bound fraction passes the column unretained and is measured downstream by means of a conventional HPLC detector. The theoretical model presented here relates the detector response to the most important instrumental and biochemical parameters such as dispersion, reaction time, concentration and affinity of the affinity protein and the number of binding sites. The theoretical concept is validated using fluorescein-labelled streptavidin and biotin as model system.

References

Jan 1, 1990·Methods in Enzymology·N M Green
Feb 18, 1994·Journal of Chromatography. B, Biomedical Applications·A J OosterkampJ van de Greef
May 1, 1993·Analytical Chemistry·P C Gunaratna, G S Wilson

❮ Previous
Next ❯

Citations

Nov 26, 2010·Analytical and Bioanalytical Chemistry·Jeroen KoolWilfried M A Niessen
Apr 11, 2012·International Journal of Molecular Sciences·Christiaan J MalherbeElizabeth Joubert
Oct 29, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Lutea A A de JongRainer Bischoff

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.