Inexpensive and relatively quick to produce Homogenous – a single clone of an antibody with a single affinity and immunoglobulin subclassįar fewer host species, limited to just mouse, rat and rabbit Heterogenous – contains a mixture of antibodies with varying affinities and immunogloblulin subclasses Monoclonal antibodies vs Polyclonal antibodies: Features of Monoclonal antibodies compared to Polyclonal Antibodies Polyclonal This is very useful in drug detection when many possible combinations of the drug are to be tested in a patient. For example, tricyclic anti-depressants, which have a similar overall structure with substitutions of differing atoms into the cyclic structure. sensitivity requirements and cross reactivity levels can be specified and monoclonal antibodies screened to identify any cell lines exhibiting the required characteristics. Monoclonals can also be generated to cross-react with groups of molecules. Specific characteristics of an antibody can be identified and selected, e.g. This hybridoma cell line, once stabilized via single cell cloning, can be frozen and stored under liquid nitrogen, allowing the antibody to be produced in vitro, in large quantities when required. Monoclonal antibodies can be raised against many targets. Meaning that not only do they produce a single antibody against a specific epitope, but they can also replicate indefinitely.Īn excellent background article into the research of monoclonal antibodies and their discovery by Köhler and Milstein has been produced by – the article can be found here. The resulting cell type is called a hybridoma. Hybridomas have the characteristics of both B-cells and myelomas. In 1975 Georges Jean Franz Köhler and César Milstein developed a technology to fuse mouse myleoma cells with B-cells. This phenomenon is known as the Hayflick limit, named after Leonard Hayflick after his work in 1961. This is a problem if one wishes to have a long-term source of a specific monoclonal antibody, produced by only one B-cell clone. The antibody producing B-cells are isolated from the spleen and lymph nodes of the immunized animals and grown, or cultured, in the laboratory. However, these cells are not immortal, replicating only 50 – 70 times before undergoing senescence and ceasing to produce antibodies. Monoclonal antibodies are created with a single B-cell clone from one animal, generating antibodies to a single specific epitope. The first stage in monoclonal antibody production is through immunization of an animal – which, for a number of reasons, tend to be mice. The Monoclonal Antibody Production Process This process – multiple immunizations followed by affinity purification – ultimately lends itself to the production of high titer, high-affinity polyclonal antibodies against the antigen of interest. This is then typically enriched using affinity purification. Following this, the serum containing the desired antibodies is collected. This is usually followed by further immunizations to boost the levels of circulating antibodies – the titer. To do this, the animals are typically immunized with a specific antigen to elicit an immune response. Polyclonal antibodies can be raised in many different species, such as goats, sheep, or rabbits. The result is the production large numbers of antibodies, each with different specificities and epitope affinities, known as ‘polyclonal antibodies’. In nature, the primary immune response to a foreign antigen involves the activation of large numbers of B-cells to protect the body.Įach B-cell targets a single antigen, and produces antibodies against a specific epitope of that antigen. So what are the difference in their production? And how does this affect their various features? The Polyclonal Antibody Production Process However, the very nature of how they are produced can often make one more suitable for a given application over another. Monoclonal Antibodies vs Polyclonal Antibodies: what are the differences and how they will affect your experiments Monoclonal antibodies vs Polyclonal antibodies: The different production processesīoth polyclonal and monoclonal antibodies share a huge number of applications, such as IHC and WB, in common.
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