A radioimmunoassay (RIA) is an

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 immunoass ...

that uses

radiolabeled A radioactive tracer, radiotracer, or radioactive label is a chemical compound in which one or more atoms have been replaced by a radionuclide so by virtue of its radioactive decay it can be used to explore the mechanism of chemical reactions by tr ...

molecules in a stepwise formation of

immune complex An immune complex, sometimes called an antigen-antibody complex or antigen-bound antibody, is a molecule formed from the binding of multiple antigens to antibodies. The bound antigen and antibody act as a unitary object, effectively an antigen o ...

es. A RIA is a very sensitive

in vitro ''In vitro'' (meaning in glass, or ''in the glass'') studies are performed with microorganisms, cells, or biological molecules outside their normal biological context. Colloquially called " test-tube experiments", these studies in biology ...

assay technique used to measure concentrations of substances, usually measuring

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 respons ...

concentrations (for example,

hormone A hormone (from the Greek participle , "setting in motion") is a class of signaling molecules in multicellular organisms that are sent to distant organs by complex biological processes to regulate physiology and behavior. Hormones are require ...

levels in

blood Blood is a body fluid in the circulatory system of humans and other vertebrates that delivers necessary substances such as nutrients and oxygen to the cells, and transports metabolic waste products away from those same cells. Blood in the cir ...

) by use of antibodies. Although the RIA technique is extremely sensitive and extremely

specific Specific may refer to: * Specificity (disambiguation) * Specific, a cure or therapy for a specific illness Law * Specific deterrence, focussed on an individual * Specific finding, intermediate verdict used by a jury in determining the fina ...

, 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 Immunoradiometric assay (IRMA) is an assay that uses radiolabeled antibodies. It differs from conventional radioimmunoassay (RIA) in that the compound to be measured combines immediately with the radiolabeled antibodies, rather than displacing anot ...

(IRMA) is an immunoassay that uses radiolabeled molecules but in an immediate rather than stepwise way. A

radioallergosorbent test A radioallergosorbent test (RAST) is a blood test using radioimmunoassay test to detect specific IgE antibodies in order to determine the substances a subject is allergic to. This is different from a skin allergy test, which determines allergy by ...

(RAST) is an example of radioimmunoassay. It is used to detect the causative

allergen An allergen is a type of antigen that produces an abnormally vigorous immune response in which the immune system fights off a perceived threat that would otherwise be harmless to the body. Such reactions are called allergies. In technical terms ...

for an

allergy Allergies, also known as allergic diseases, refer a number of conditions caused by the hypersensitivity of the immune system to typically harmless substances in the environment. These diseases include hay fever, food allergies, atopic derm ...

Method

Classically, to perform a radioimmunoassay, a known quantity of an

is made

radioactive Radioactive decay (also known as nuclear decay, radioactivity, radioactive disintegration, or nuclear disintegration) is the process by which an unstable atomic nucleus loses energy by radiation. A material containing unstable nuclei is consi ...

, frequently by labeling it with gamma-radioactive

isotopes of 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 yea ...

, such as 125-I, attached to

tyrosine -Tyrosine or tyrosine (symbol Tyr or Y) or 4-hydroxyphenylalanine is one of the 20 standard amino acids that are used by cells to synthesize proteins. It is a non-essential amino acid with a polar side group. The word "tyrosine" is from the G ...

. This radiolabeled antigen is then mixed with a known amount of

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 ...

for that antigen, and as a result, the two specifically bind to one another. Then, a sample of serum from a patient containing an unknown quantity of that same antigen is added. This causes the unlabeled (or "cold") antigen from the serum to compete with the radiolabeled antigen ("hot") for antibody binding sites. As the

concentration In chemistry, concentration is the abundance of a constituent divided by the total volume of a mixture. Several types of mathematical description can be distinguished: '' mass concentration'', ''molar concentration'', '' number concentration'', ...

of "cold" antigen is increased, more of it binds to the antibody, displacing the radiolabeled variant, and reducing the ratio of antibody-bound radiolabeled antigen to free radiolabeled antigen. The bound antigens are then separated and the radioactivity of the free(unbound) antigen remaining in the

supernatant In an aqueous solution, precipitation is the process of transforming a dissolved substance into an insoluble solid from a super-saturated solution. The solid formed is called the precipitate. In case of an inorganic chemical reaction leading ...

is measured using a

gamma counter A gamma counter is an instrument to measure gamma radiation emitted by a radionuclide. Unlike survey meters, gamma counters are designed to measure small samples of radioactive material, typically with automated measurement and movement of multi ...

. This method can be used for any biological molecule in principle and is not restricted to serum antigens, nor is it required to use the indirect method of measuring the free antigen instead of directly measuring the captured antigen. For example, if it is undesirable or not possible to radiolabel the antigen or target molecule of interest, a RIA can be done if two different antibodies that recognize the target are available and the target is large enough (e.g., a protein) to present multiple

epitope An epitope, also known as antigenic determinant, is the part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells. The epitope is the specific piece of the antigen to which an antibody binds. The p ...

s to the antibodies. One antibody would be radiolabeled as above while the other would remain unmodified. The RIA would begin with the "cold" unlabeled antibody being allowed to interact and bind to the target molecule in solution. Preferably, this unlabeled antibody is immobilized in some way, such as coupled to an

agarose Agarose is a heteropolysaccharide, generally extracted from certain red seaweed. It is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose. Agarose is ...

bead, coated to a surface, etc. Next, the "hot" radiolabeled antibody is allowed to interact with the first antibody-target molecule complex. After extensive washing, the direct amount of radioactive antibody bound is measured and the amount of target molecule quantified by comparing it to a reference amount assayed at the same time. This method is similar in principle to the non-radioactive sandwich

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 presen ...

method.

History

This method was developed by

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 "Biographica ...

and

Rosalyn Sussman 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 ...

at the

Veterans Administration Hospital Veterans' health care in the United States is separated geographically into 19 regions (numbered 1, 2, 4-10, 12 and 15–23) In January 2002, the Veterans Health Administration announced the merger of VISNs 13 and 14 to create a new, combined netw ...

the Bronx The Bronx () is a borough of New York City, coextensive with Bronx County, in the state of New York. It is south of Westchester County; north and east of the New York City borough of Manhattan, across the Harlem River; and north of the New Y ...

, New York. This revolutionary development earned Dr. Yalow the Nobel Prize for Medicine in 1977, the second woman ever to win it. In her acceptance speech, Dr. Yalow said, "The world cannot afford the loss of the talents of half its people if we are to solve the many problems which beset us." Yalow shared the Nobel Prize with

Roger Guillemin Roger Charles Louis Guillemin (born January 11, 1924) is a French-American neuroscientist. He received the National Medal of Science in 1976, and the Nobel prize for medicine in 1977 for his work on neurohormones, sharing the prize that year ...

, and

Andrew Schally Andrzej Viktor "Andrew" Schally (born 30 November 1926) is an American endocrinologistAndrew V. Schally"Andrew V. Schally" ''Encyclopædia Britannica''. of Polish ancestry, who was a corecipient, with Roger Guillemin and Rosalyn Sussman Yalow, of ...

who earned the prize based on their research into "the peptide hormone production of the brain".

References

steps i
radioimmunoassay
technique

External links

*

{{Immunologic techniques and tests Endocrine procedures Immunologic tests Radiobiology