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Hybridoma
Hybridoma technology is a method for producing large quantities of monoclonal antibodies by fusing antibody producing B cells with myeloma cells (cancerous B cells). This creates hybrid cells, ''hybridomas,'' that produce the antibody from their parent B cell whilst maintaining the properties of the parental myeloma cell line being immortal (endlessly reproducing) and having desirable properties for cell culture. The B cells to be used are generally gathered from animals who have been immunized with an antigen against which an antibody targeting it is desired. After forming hybridomas any non-hybrid cells are killed before screening and monoclonalization to create hybridoma lines that are derived from one parental cell and thus producing the same antibody against the desired target. The production of monoclonal antibodies was invented by César Milstein and Georges J. F. Köhler in 1975. They shared the Nobel Prize of 1984 for Medicine and Physiology with Niels Kaj Jerne, ...
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Hybridoma Cells Grown In Tissue Culture
Hybridoma technology is a method for producing large quantities of monoclonal antibodies by fusing antibody producing B cell, B cells with Multiple myeloma, myeloma cells (cancerous B cells). This creates hybrid cells, ''hybridomas,'' that produce the antibody from their parent B cell whilst maintaining the properties of the parental myeloma cell line being Immortalised cell line, immortal (endlessly reproducing) and having desirable properties for cell culture. The B cells to be used are generally gathered from animals who have been immunized with an antigen against which an antibody targeting it is desired. After forming hybridomas any non-hybrid cells are killed before screening and Monoclonality, monoclonalization to create hybridoma lines that are derived from one parental cell and thus producing the same antibody against the desired target. The production of monoclonal antibodies was invented by César Milstein and Georges J. F. Köhler in 1975. They shared the Nobel Pri ...
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Hybridoma Technology
Hybridoma technology is a method for producing large quantities of monoclonal antibodies by fusing antibody producing B cells with myeloma cells (cancerous B cells). This creates hybrid cells, ''hybridomas,'' that produce the antibody from their parent B cell whilst maintaining the properties of the parental myeloma cell line being immortal (endlessly reproducing) and having desirable properties for cell culture. The B cells to be used are generally gathered from animals who have been immunized with an antigen against which an antibody targeting it is desired. After forming hybridomas any non-hybrid cells are killed before screening and monoclonalization to create hybridoma lines that are derived from one parental cell and thus producing the same antibody against the desired target. The production of monoclonal antibodies was invented by César Milstein and Georges J. F. Köhler in 1975. They shared the Nobel Prize of 1984 for Medicine and Physiology with Niels Kaj Jerne, wh ...
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Monoclonals
A monoclonal antibody (mAb, more rarely called moAb) is an antibody produced from a Lineage (evolution), cell lineage made by cloning a unique white blood cell. All subsequent antibodies derived this way trace back to a unique parent cell. Monoclonal antibodies are identical and can thus have Valence (chemistry), monovalent affinity, binding only to a particular epitope (the part of an antigen that is recognized by the antibody). In contrast, polyclonal antibodies are mixtures of antibodies derived from multiple plasma cell lineages which each bind to their particular target epitope. Artificial antibodies known as bispecific monoclonal antibodies can also be engineered which include two different antigen binding sites (Fragment antigen-binding region, FABs) on the same antibody. It is possible to produce monoclonal antibodies that specifically bind to almost any suitable substance; they can then serve to detect or purify it. This capability has become an investigative tool in b ...
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César Milstein
César Milstein, CH, FRS (8 October 1927 – 24 March 2002) was an Argentine biochemist in the field of antibody research. Milstein shared the Nobel Prize in Physiology or Medicine in 1984 with Niels Kaj Jerne and Georges J. F. Köhler for developing the hybridoma technique for the production of monoclonal antibodies. Biography Milstein was born in Bahía Blanca, Argentina. His parents were Máxima (Vanarks) and Lázaro Milstein, a Jewish Ukrainian immigrant. He graduated from the University of Buenos Aires and obtained a PhD under Professor Stopani (Professor of Biochemistry). Later he became a member of the Medical Research Council Laboratory of Molecular Biology, Cambridge, England; he acquired British citizenship and had dual British-Argentinian nationality. In 1956, he received an award from the Sociedad Argentina de Investigation eon Bio Quimica (SAIB) for his work on enzyme kinetics with the enzyme aldehyde dehydrogenase. In 1958, funded by the British Council, he j ...
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Antibody
An antibody (Ab) or immunoglobulin (Ig) is a large, Y-shaped protein belonging to the immunoglobulin superfamily which is used by the immune system to identify and neutralize antigens such as pathogenic bacteria, bacteria and viruses, including those that cause disease. Each individual antibody recognizes one or more specific antigens, and antigens of virtually any size and chemical composition can be recognized. Antigen literally means "antibody generator", as it is the presence of an antigen that drives the formation of an antigen-specific antibody. Each of the branching chains comprising the "Y" of an antibody contains a paratope that specifically binds to one particular epitope on an antigen, allowing the two molecules to bind together with precision. Using this mechanism, antibodies can effectively "tag" the antigen (or a microbe or an infected cell bearing such an antigen) for attack by cells of the immune system, or can neutralize it directly (for example, by blocking a p ...
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Cell Culture
Cell culture or tissue culture is the process by which cell (biology), cells are grown under controlled conditions, generally outside of their natural environment. After cells of interest have been Cell isolation, isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich growth medium, medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (Carbon dioxide, CO2, Oxygen, O2), and regulates the physio-chemical environment (Buffer solution, pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as a suspension culture. T ...
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Monoclonality
In biology, monoclonality refers to the state of a line of cells that have been derived from a single clonal origin. Thus, "monoclonal cells" can be said to form a single clone. The term ''monoclonal'' comes . The process of replication can occur ''in vivo'', or may be stimulated ''in vitro'' for laboratory manipulations. The use of the term typically implies that there is some method to distinguish between the cells of the original population from which the single ancestral cell is derived, such as a random genetic alteration, which is inherited by the progeny. Common usages of this term include: * Monoclonal antibody: a single hybridoma cell, which by chance includes the appropriate V(D)J recombination to produce the desired antibody, is cloned to produce a large population of identical cells. In informal laboratory jargon, the monoclonal antibodies isolated from cell culture supernatants of these hybridoma clones (hybridoma lines) are simply called ''monoclonals''. * Mono ...
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Leonard Herzenberg
Leonard Arthur "Len" Herzenberg (November 5, 1931 – October 27, 2013) was an immunologist, geneticist and professor at Stanford University. His contributions to the development of cell biology made it possible to sort viable cells by their specific properties. Education Herzenberg was born in New York City, on November 5, 1931. He received his bachelor's degree in 1952 from Brooklyn College in biology and chemistry. In 1955, he received his Ph.D. from California Institute of Technology in biochemistry with a specialization in immunology for studies on cytochrome in Neurospora. Career After school he was a postdoctoral fellow at the American Cancer Society, working in France at the Pasteur Institute. He returned to the United States in 1957 and worked for the National Institutes of Health as an officer in the Public Health Service department. He started working at Stanford in 1959. He eventually earned the title Professor of Genetics. In 1970 Herzenberg developed the fluor ...
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Western Blot
The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. Besides detecting the proteins, this technique is also utilized to visualize, distinguish, and quantify the different proteins in a complicated protein combination. Western blot technique uses three elements to achieve its task of separating a specific protein from a complex: separation by size, transfer of protein to a solid support, and marking target protein using a primary and secondary antibody to visualize. A synthetic or animal-derived antibody (known as the primary antibody) is created that recognizes and binds to a specific target protein. The electrophoresis membrane is washed in a solution containing the primary antibody, before excess antibody is washed off. A secondary antibody is added which recognizes and binds to the primary antibod ...
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Aminopterin
Aminopterin (or 4-aminopteroic acid), the 4–amino derivative of folic acid, is an antineoplastic drug with immunosuppressive properties often used in chemotherapy. Aminopterin is a synthetic derivative of pterin. Aminopterin works as an enzyme inhibitor by competing for the folate binding site of the enzyme dihydrofolate reductase. Its binding affinity for dihydrofolate reductase effectively blocks tetrahydrofolate synthesis. This results in the depletion of nucleotide precursors and inhibition of DNA, RNA, and protein synthesis. It is classified as an extremely hazardous substance in the United States as defined in Section 302 of the U.S. Emergency Planning and Community Right-to-Know Act (42 U.S.C. 11002), and is subject to strict reporting requirements by facilities which produce, store, or use it in significant quantities. Uses Discovered by Yellapragada Subbarow, the drug was first used by Sidney Farber in 1947 to induce remissions among children with leukemia. Amino ...
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