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Production and characteristics of Monoclonal antibodies:-
#1
Antibodies are actually the proteins which are produced by B-lymphocytes present in the immune system. They are produced in response to the foreign object which enters the body called as antigens. Their actual function is to bind with the antigen and make it prominent to the immune system so that it can be destroyed. Antibodies act as markers and the site where they bind to the antigen is called as epitope. Epitope is a small amino acid sequence which can easily be recognized by the antibody. Antibody has the ability to bind to a particular epitope and it is not necessary that whole antigen is present during the attachment of antibody to the epitope.

How are monoclonal antibodies produced:-
As monoclonal antibodies are produced in the laboratory that is why there is a certain procedure for their production. General procedure of production is as follows;

1) Myeloma cells, which are produced due to the malfunctioning of body cells and become cancerous cells, are fused with the spleen cells taken from mouse produced in the laboratory. Methods like electroporation are used to fuse these two cells.

2) The substance produced as a result of fusion of two cells is celled as Hybridoma.

3) In the second step, a medium is prepared for Hybridoma. In this medium, there are two types of cells that are hypoxanthine-guanine phosphoribosyl transferase which are the myeloma cells and B cells are hypoxanthine-guanine phosphoribosyl transferase plus. When in the culture, myeloma cells will combine with other myeloma cells or B cells will combine with other B cells, they will not be able to survive. When myeloma cells will combine with B cells, only those cells will survive in the medium.

4) Now the combination of two cells that is myeloma and B cells is cloned and then diluted. The resulting substance will be the antibodies. Scientists will check, if they have the ability to bind to the specific antigen or not.

5) Different tests are performed to see the binding of antibody to the antigen for example ELISA or Antigen microarray Assay. Best clone is saved for future purposes.

6) When the monoclonal antibody is produced, there are some impurities also in the medium which are in the form of hormones, enzymes and growth factors. Filtration method is used to remove some of the lipids, cell debris and other useless materials from the medium. Antibodies are taken out of the culture by precipitating the medium with sodium sulphate or ammonium sulphate. Antibodies are able to precipitate out even at low concentrations of salts. Purity of monoclonal antibodies is checked with ELISA kits.

Characteristics of monoclonal antibodies:-
Monoclonal antibodies have the ability to bind to a particular antigen and attack it. Once an antibody is activated against some diseases especially the diseases of childhood like chicken pox and measles, then it keeps on creating resistance against that particular disease until the disease is fully removed from the body. Antibodies have enabled the scientists to develop vaccines which also act against a particular disease or antigen.
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#2
Can you explain why Pig IgG is present in the ELISA sensitizing solution in addition to the antibody specific for the ELISA kit? Thank you.
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#3
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Production and characteristics of Monoclonal antibodies:-00