MoAb are a perpetual source of antibodies with consistent specificity and affinity. MoAb were discovered by Kohler and Milstein. The solution to these drawbacks of PoAb was found with the discovery of monoclonal antibodies (MoAb).
Animals have to be given booster injections each time before collecting PoAb. The major limitations in using PoAb are the huge variations in affinity and specificity of PoAb from lot-to-lot. PoAb were used to protect against infections like diphtheria as early as 1894. PoAb have various applications of analytical, diagnostic, and therapeutic significance. Polyclonal antibodies (PoAb) can be raised by immunizing animals against the antigen of choice. Antibodies are produced by B-lymphocytes after differentiation of B-lymphocytes into plasma cells. Antibodies are produced either to neutralize or to eliminate antigens or pathogens. Ashish Swarup Verma, in Animal Biotechnology, 2014 What You can Expect to KnowĪntibody production is a hallmark of the adaptive immune response. An unknown serum is tested for its ability to bind to any of the proteins on the strip, and the reaction is “developed” with a labeled antisera against immunoglobulin, as for the ELISA.Īnchal Singh. Proteins from a viral lysate are separated, often using polyacrylamide gel electrophoresis, and crosslinked to a cellulose strip. In contrast to the ELISA, the Western blot is a qualitative test that provides information about the specificity of the test antibody.
An antigen, either whole virus, a viral protein, or a viral peptide, is bound to a substrate, and then incubated with serial dilutions of test antibody adherence of the test antibody is determined with a conjugated antiserum directed against immunoglobulin of the species under test. Of these, the most commonly used is the ELISA assay, which can readily be adapted to quantitation, automation, and rapid throughput. There are many alternative assays that measure the ability of the antibody to bind to viral antigens, including hemagglutination inhibition, immunofluorescence, Western blot, and ELISA (enzyme linked immunosorbent assay). However, neutralization tests cannot be used for some important viruses, such as hepatitis B and C viruses, that cannot readily be grown in cell culture. One common technique involves the use of a single viral inoculum, such as 100 PFU (plaque forming units) serial dilutions of a test antibody are tested to determine the highest dilution that will reduce the plaque count by 50%. This test depends on the availability of a convenient method to measure viral infectivity, often a plaque assay. The canonical assay is the neutralization test, in which antibody is tested for its ability to reduce viral infectivity.
There are many methods to measure antibody to viruses, and both the kinetics of the response and its biological significance depend upon the assay used. John Wherry, David Masopust, in Viral Pathogenesis (Third Edition), 2016 2.1 Measures of Antibody While FCA remains overall the most effective adjuvant for polyclonal antibody production, the extensive histologic lesions it produces will continue to stimulate the development of alternative adjuvants producing less tissue destruction and potential pain and distress.Į. The choice of adjuvant and immunization schedules are critical components of the polyclonal antibody production process that are frequently overlooked as researchers resort to standard published methodologies ( Cooper and Paterson, 2008, 2009 Harlow, 1988) that may or may not be applicable to their needs. The schedules and methodologies used to immunize rabbits and produce polyclonal antibodies will continue to vary dependent upon the immunogen, the adjuvant, and the end purpose of the antibody. Polyclonal antibody production will remain an essential research activity and rabbits will continue to serve as one of the primary species used in polyclonal antibody production. Stills, in The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents, 2012 Summary