![]() With affinity purification of serum against small antigen targets, the advantages of polyclonal antibodies are further extended. For general research applications, however, the advantages of polyclonal antibodies typically outweigh the few advantages that monoclonal antibodies provide. This is typically cost and time prohibitive.įor applications such as diagnostic manufacturing and therapeutic drug development that require large volumes of identical antibody specific to a single epitope, monoclonal antibodies can be an ideal solution. To offset many of these disadvantages, it is necessary to produce a pool of several monoclonal antibodies.More susceptible to binding changes when labeled.More sensitive to pH and buffer conditions. ![]() Less ideal for application requiring quick capture of the target protein.Less robust for detecting the protein in a denatured state or altered conformation.Cell culture and purification capabilities required.More demanding storage conditions for the clone.Small changes in the epitope’s structure often render the monoclonal antibody unable to detect the target protein.Requires significantly more time to produce and develop the hybridized clone.Significantly more expensive to produce.Can provide better results in assays requiring quantification of the protein levels.Can produce large quantities of identical antibody.Each individual antibody in a polyclonal mixture is technically a monoclonal antibody however, this term generally refers to a process by which the actual B-cell is isolated and fused to an immortal hybridoma cell line so that large quantities of identical antibody can be generated. Affinity purification of the serum will typically be required to minimize cross reactivityĪ Monoclonal antibody, by contrast, represents antibody from a single antibody producing B cell and therefore only binds with one unique epitope.Higher potential for cross reactivity due to recognizing multiple epitopes.Variability between different batches produced in different animals at different times.Less likely to affect binding capability. Much easier to couple with antibody labels.Superior for use in detecting a native protein in multiple assay types.Significantly more robust when assaying proteins that show slight variations in individual epitopes such as denaturation, polymorphism or conformational changes.Ideal in assays requiring quick capture of the protein such as IP or ChIP. Superior antibody affinity generally results in quicker binding to target antigen.Greater ability to quickly capture the target protein. Ideal as the capture antibody in a Sandwich ELISA.Offers greater sensitivity for detecting proteins that are present in low quantities in a sample since multiple antibodies will bind to multiple epitopes on the protein.In general, ability to detect multiple epitopes gives more robust detection.Higher overall antibody affinity against the antigen due to recognition of multiple epitopes.Highly stable and tolerant of pH or buffer changes.Purified antibody ready to use in under four months. Each of these individual antibodies recognizes a unique epitope that is located on that antigen. Recommendations are provided for optimizing the immunization response.A Polyclonal Antibody represents a collection of antibodies from different B cells that recognize multiple epitopes on the same antigen. In this article, the critical steps in the production of polyclonal and monoclonal antibodies are described, specifically including selection of the animal species and its age, injection protocol, and ascites tapping. However, by carefully designing an immunization protocol and by optimizing the immunization response, it is possible to minimize animals' pain and distress while obtaining optimal immune responses. ![]() Antibody production requires a substantial number of animals, and the animals are subjected to a number of invasive procedures such as antigen injection and blood collection. Both products have become essential instruments in fundamental immunological research, immunohistochemistry, diagnostic testing, and vaccine quality control. Antibodies include those secreted by a single clone of B lymphocytes, termed monoclonal antibodies, and those produced by a mixture of various B lymphocyte clones, termed polyclonal antibodies. Antibodies are valuable tools in the laboratory and clinic.
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