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Immunoassay
An immunoassay (IA) is a biochemical test that measures the presence or concentration of a macromolecule or a small molecule in a solution through the use of an antibody (usually) or an antigen (sometimes). The molecule detected by the immunoassay is often referred to as an "analyte" and is in many cases a protein, 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 serum or urine are frequently measured using immunoassays for medical and research purposes. 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 can be carried out simply by mixing the reagents and sample and making a p ...
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Radioimmunoassay
A radioimmunoassay (RIA) is an immunoassay that uses radiolabeled molecules in a stepwise formation of immune complexes. A RIA is a very sensitive in vitro assay technique used to measure concentrations of substances, usually measuring antigen concentrations (for example, hormone levels in blood) by use of antibodies. Although the RIA technique is extremely sensitive and extremely specific, requiring specialized equipment, it remains among the least expensive methods to perform such measurements. It requires special precautions and licensing, since radioactive substances are used. In contrast, an immunoradiometric assay (IRMA) is an immunoassay that uses radiolabeled molecules but in an immediate rather than stepwise way. A radioallergosorbent test (RAST) is an example of radioimmunoassay. It is used to detect the causative allergen for an allergy. Method Classically, to perform a radioimmunoassay, a known quantity of an antigen is made radioactive, frequently by labeling it ...
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ELISA
The enzyme-linked immunosorbent assay (ELISA) (, ) is a commonly used analytical biochemistry assay, first described by Eva Engvall and Peter Perlmann in 1971. The assay uses a solid-phase type of enzyme immunoassay (EIA) to detect the presence of a ligand (commonly a protein) in a liquid sample using antibodies directed against the protein to be measured. ELISA has been used as a diagnostic tool in medicine, plant pathology, and biotechnology, as well as a quality control check in various industries. In the most simple form of an ELISA, antigens from the sample to be tested are attached to a surface. Then, a matching antibody is applied over the surface so it can bind the antigen. This antibody is linked to an enzyme and then any unbound antibodies are removed. In the final step, a substance containing the enzyme's substrate is added. If there was binding, the subsequent reaction produces a detectable signal, most commonly a color change. Performing an ELISA involves at le ...
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Rosalyn Yalow
Rosalyn Sussman Yalow (July 19, 1921 – May 30, 2011) was an American medical physicist, and a co-winner of the 1977 Nobel Prize in Physiology or Medicine (together with Roger Guillemin and Andrew Schally) for development of the radioimmunoassay technique. She was the second woman (after Gerty Cori), and the first American-born woman, to be awarded the Nobel Prize in Physiology or Medicine. Biography Childhood Rosalyn Sussman Yalow was born in the Bronx, New York, the daughter of Clara (née Zipper) and Simon Sussman, and was raised in a Jewish household. She went to Walton High School (Bronx), New York City. After high school, she attended the all-female, tuition-free Hunter College, where her mother hoped she would learn to become a teacher. Instead, Yalow decided to study physics. College Yalow knew how to type, and was able to get a part-time position as a secretary to Dr. Rudolf Schoenheimer, a leading biochemist at Columbia University's College of Physicians and Surgeon ...
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Enzyme Multiplied Immunoassay Technique
Enzyme multiplied immunoassay technique (EMIT) is a common method for qualitative and quantitative determination of therapeutic and recreational drugs and certain proteins in serum and urine. It is an immunoassay in which a drug or metabolite in the sample competes with a drug/metabolite labelled with an enzyme, to bind to an antibody. The more drug there is in the sample, the more free enzyme there will be, and the increased enzyme activity causes a change in color. Determination of drug levels in serum is particularly important when the difference in the concentrations needed to produce a therapeutic effect and adverse side reactions (the therapeutic window) is small. EMIT therapeutic drug monitoring tests provide accurate information about the concentration of such drugs such as immunosuppressant drugs and some antibiotics. EMIT urine assays for drugs such as cannabinoids, morphine, and amphetamine are designed to detect the drug itself or a metabolite of the drug present in a ...
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Solomon Berson
Solomon Aaron Berson (April 22, 1918 – April 11, 1972) was an American physician and scientist whose discoveries, mostly together with Rosalyn Yalow, caused major advances in clinical biochemistry.Rall JE. ''Solomon A. Berson''. In "Biographical Memoirs". National Academy of Sciences 1990;59:54-71. Fulltext Five years after Berson's death, Yalow received a Nobel Prize, which cannot be awarded posthumously, for their joint work on the radioimmunoassay. Biography Early life Born in New York City, Berson was a keen musician and chess player. He graduated from the City College of New York in 1938. After failing to obtain a place in medical school he earned an MSc (1939) and an anatomy instructorship at New York University before finally securing a place in NYU medical school in 1941. He completed his degree (Alpha Omega Alpha) in 1945, and after internships in Boston and two years in the army he returned to New York to do an internal medicine residency at the Bronx Veterans Ad ...
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Antibody
An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein used by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of the pathogen, called an antigen. Each tip of the "Y" of an antibody contains a paratope (analogous to a lock) that is specific for one particular epitope (analogous to a key) on an antigen, allowing these two structures to bind together with precision. Using this binding mechanism, an antibody can ''tag'' a microbe or an infected cell for attack by other parts of the immune system, or can neutralize it directly (for example, by blocking a part of a virus that is essential for its invasion). To allow the immune system to recognize millions of different antigens, the antigen-binding sites at both tips of the antibody come in an equally wide variety. In contrast, the remainder of the antibody is relatively constant. It only occurs in a few vari ...
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Horseradish Peroxidase
The enzyme horseradish peroxidase (HRP), found in the roots of horseradish, is used extensively in biochemistry applications. It is a metalloenzyme with many isoforms, of which the most studied type is C. It catalyzes the oxidation of various organic substrates by hydrogen peroxide. Structure The structure of the enzyme was first solved by X-ray crystallography in 1997; and has since been solved several times with various substrates. It is a large alpha-helical glycoprotein which binds heme as a redox cofactor. Substrates Alone, the HRP enzyme, or conjugates thereof, is of little value; its presence must be made visible using a substrate that, when oxidized by HRP using hydrogen peroxide as the oxidizing agent, yields a characteristic color change that is detectable by spectrophotometric methods. Numerous substrates for horseradish peroxidase have been described and commercialized to exploit the desirable features of HRP. These substrates fall into several distinct ...
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Antigen
In immunology, an antigen (Ag) is a molecule or molecular structure or any foreign particulate matter or a pollen grain that can bind to a specific antibody or T-cell receptor. The presence of antigens in the body may trigger an immune response. The term ''antigen'' originally referred to a substance that is an antibody generator. Antigens can be proteins, peptides (amino acid chains), polysaccharides (chains of monosaccharides/simple sugars), lipids, or nucleic acids. Antigens are recognized by antigen receptors, including antibodies and T-cell receptors. Diverse antigen receptors are made by cells of the immune system so that each cell has a specificity for a single antigen. Upon exposure to an antigen, only the lymphocytes that recognize that antigen are activated and expanded, a process known as clonal selection. In most cases, an antibody can only react to and bind one specific antigen; in some instances, however, antibodies may cross-react and bind more than one antigen ...
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Analyte
An analyte, component (in clinical chemistry), or chemical species is a substance or chemical constituent that is of interest in an analytical procedure. The purest substances are referred to as analytes, such as 24 karat gold, NaCl, water, etc. In reality, no substance has been found to be 100% pure in its quality, so a substance that is found to be most pure (for some metals, 99% after electrolysis) is called an analyte. See also *Analytical chemistry *Immunoassay *Magnetic immunoassay Magnetic immunoassay (MIA) is a type of diagnostic immunoassay using magnetic beads as labels in lieu of conventional enzymes (ELISA), radioisotopes (RIA) or fluorescent moieties ( fluorescent immunoassays) to detect a specified analyte. MIA involv ... References Analytical chemistry {{Analytical-chemistry-stub ...
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Alkaline Phosphatase
The enzyme alkaline phosphatase (EC 3.1.3.1, alkaline phosphomonoesterase; phosphomonoesterase; glycerophosphatase; alkaline phosphohydrolase; alkaline phenyl phosphatase; orthophosphoric-monoester phosphohydrolase (alkaline optimum), systematic name phosphate-monoester phosphohydrolase (alkaline optimum)) catalyses the following reaction: : a phosphate monoester + H2O = an alcohol + phosphate Alkaline phosphatase has the physiological role of dephosphorylating compounds. The enzyme is found across a multitude of organisms, prokaryotes and eukaryotes alike, with the same general function but in different structural forms suitable to the environment they function in. Alkaline phosphatase is found in the periplasmic space of '' E. coli'' bacteria. This enzyme is heat stable and has its maximum activity at high pH. In humans, it is found in many forms depending on its origin within the body – it plays an integral role in metabolism within the liver and development wit ...
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Radioactive Iodine
There are 37 known isotopes of iodine (53I) from 108I to 144I; all undergo radioactive decay except 127I, which is stable. Iodine is thus a monoisotopic element. Its longest-lived radioactive isotope, 129I, has a half-life of 15.7 million years, which is far too short for it to exist as a primordial nuclide. Cosmogenic sources of 129I produce very tiny quantities of it that are too small to affect atomic weight measurements; iodine is thus also a mononuclidic element—one that is found in nature only as a single nuclide. Most 129I derived radioactivity on Earth is man-made, an unwanted long-lived byproduct of early nuclear tests and nuclear fission accidents. All other iodine radioisotopes have half-lives less than 60 days, and four of these are used as tracers and therapeutic agents in medicine. These are 123I, 124I, 125I, and 131I. All industrial production of radioactive iodine isotopes involves these four useful radionuclides. The isotope 135I has a half-life less than se ...
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Glucose Oxidase
The glucose oxidase enzyme (GOx or GOD) also known as notatin (EC number 1.1.3.4) is an oxidoreductase that catalyses the oxidation of glucose to hydrogen peroxide and D-glucono-δ-lactone. This enzyme is produced by certain species of fungi and insects and displays antibacterial activity when oxygen and glucose are present. Glucose oxidase is widely used for the determination of free glucose in body fluids (medical testing), in vegetal raw material, and in the food industry. It also has many applications in biotechnologies, typically enzyme assays for biochemistry including biosensors in nanotechnologies. It was first isolated by Detlev Müller in 1928 from ''Aspergillus niger''. Function Several species of fungi and insects synthesize glucose oxidase, which produces hydrogen peroxide, which kills bacteria. Notatin, extracted from antibacterial cultures of ''Penicillium notatum'', was originally named Penicillin A, but was renamed to avoid confusion with penicillin. Notat ...
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