Monoclonal Antibodies

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Monoclonal Antibodies

Postby sciguy » Nov 11 2012 9:28 pm

Why do some antibodies would recognize both wild type and mutant forms of the protein while others would recognize only the wild type form
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Re: Monoclonal Antibodies

Postby relaxin » Nov 12 2012 2:38 am

Monoclonal antibodies recognize only one epitope. If both WT and mutant possess the same epitope, then the antibody can recognize both. If the mutant no longer has this epitope (due to truncation of the protein by frame-shift mutation or point mutation that changes a key residue on the epitope), the antibody will not be able to recognize the mutant.
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Re: Monoclonal Antibodies

Postby Steven2014 » Mar 18 2015 7:37 am

For polyclonal Antibodies, the immune response to an antigen generally involves the activation of multiple B-cells all of which target a specific epitope on that antigen. As a result a large number of antibodies are produced with different specificities and epitope affinities.

Monoclonal antibodies represent a single B lymphocyte generating antibodies to one specific epitope. B-cells can be isolated easily from the spleen and lymph nodes of immunised animals; however, these cells have a limited life span, and can only divide a limited number of times, coined the 'Hayflick limit'. This prohibits the culture of B-cells directly. For an antibody to be useful in research or industry, it must be readily available in large quantities. Due to the Hayflick limit, this would not be possible using B-cells cultured in vitro as they would eventually stop dividing and the population would die out. Consequently, in 1975 Kohler and Milstein developed a technology to fuse immortal heteromyleoma cells with lymphocytes, using poly ethylglycol (PEG) to break down cell membranes and allow mixing of the genetic material from both cell types. The resulting cell type is called a hybridoma. This hybridoma takes on the characteristics of both the lymphocyte and heteromyeloma cell, creating an immortal cell with the ability to produce antibody. As the new cell line hybridoma is a product of the fusion of one heteromyeloma cell with one B-cell, the culture only ever has one antibody within the supernatant which, when purified, is called a Monoclonal antibody.
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Re: Monoclonal Antibodies

Postby Jessica666 » Sep 18 2015 2:53 am

monoclonal antibody is stimulated by heterologous antigens and then organism produces immune reaction. It is produced by organism plasma cells. However, polyclonal antibody is cloned by one antibody which only has reaction to the epitope of a specific antigen.
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Re: Monoclonal Antibodies

Postby mdfenko » Sep 18 2015 12:43 pm

Jessica666 wrote:monoclonal antibody is stimulated by heterologous antigens and then organism produces immune reaction. It is produced by organism plasma cells. However, polyclonal antibody is cloned by one antibody which only has reaction to the epitope of a specific antigen.

you have that backwards.
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Re: Monoclonal Antibodies

Postby sile88314 » Jun 01 2016 4:00 am

relaxin wrote:Monoclonal antibodies recognize only one epitope. If both WT and mutant possess the same epitope, then the antibody can recognize both. If the mutant no longer has this epitope (due to truncation of the protein by frame-shift mutation or point mutation that changes a key residue on the epitope), the antibody will not be able to recognize the mutant.

agree with 1st floor. As supplementary, the so called "monoclonal antibody" is not absolutely monoclone, its specificity depends on the quality of the antibody production process. The more strict screening process it went through, the better specificity it would have. if you ensure the quality of your monoclonal antibody, the only reason for your antibody recognize both wild type and mutant forms of the protein would be that the antibody recognize a common epitope from both the WT and the Mutant.
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