In addition to being crucial for your normal immune response, antibodies provide powerful tools for research and diagnostic purposes. Antibodies for research and diagnostic purposes are often obtained by injecting a lab animal such as goat anti-llama biotinwith a specific antigen.
Polyclonal antibodies are produced by injecting a specific antigen into lab animals. The animal is immunized repeatedly to obtain higher tittersof the antibodies specific to the antigen. Within a few weeks, these polyclonal antibodies can be harvested and collected from the antiserum. There are several critical steps involved in the production of polyclonal antibodies. Let’s discuss some of the steps in this article.
Antigen preparation–It is important to ensure the quality and quantity of the target antigen. The production process starts with preparing the protein or peptide antigens. The specificity of the polyclonal antibody obtained depends on the priority of the antigen applied.Purification results in an increased number of specific antibodies and avoids removing many unwanted antibodies by extensive observation procedures.
Selecting the animals - When selecting the animal species, it is important to consider the amount of PAb needed, the ease of obtaining blood samples, and the phylogenetic relationship between the antigen and the animal species.A polyclonal antibody is generated in vivo by immunizing the chosen host like sheep anti-BrdU with the specific antigen or hapten- carrier conjugate. An adjuvant may be used to enhance the immune response.
Selecting and preparing adjuvant –When the antigen to which antibodies are evoked is poorly immunogenic, the immune system requires a stimulus to induce an effective immune response. An adjuvant can be used for this purpose and can direct an immune response against the most cellularDNPK1 or humoral response. Prior to the priming immunization, following the primary and each booster immunization, the animal is bled and serum prepared from the whole blood. When the antibody titer has reached an acceptable level, the production of polyclonal antibodies should be ended.
Antibody purification - In affinity purification, protein A/G affinity purification can enrich Immunoglobulin in the raw antiserum and remove the bulk of unwanted proteins. However, there is still a large quantity of non-specific IgGs in this preparation. To isolate the specific polyclonal antibodies from antiserum, antigen-specific affinity purification is often used.
Qualify control – After purification, a series of quality control tests are performed to ensure the quality of polyclonal antibodies. The absorption evaluates antibody concentration. The priority of polyclinic and the body is checked using SDS-PAGE. To conclude, new developments in vitro MAb production will gradually limit animals’ use for this purpose.
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