Antibodies are innate components of the body’s adaptive immune system that are created in response to infections known as antigens. Their major purpose is to bind to antigens via a lock-and-key mechanism, calming them. While endogenous antigens are lifesavers, their specificity limits their use outside the body.
Fortunately, breakthroughs in biotechnology and recombinant protein manufacturing have resulted in the creation of bespoke antibody approaches. Custom antibody production services allow the creation of antibodies for tasks when commercially available antibodies are insufficient.
An overview of polyclonal antibody production is provided here, covering the procedures, applications, and benefits of employing bespoke antibody services.
History of Antibodies
According to one historical version, antibodies were first mentioned in the 1890s in a book by two pioneer immunologists. The two collaborated on a report demonstrating that transferring blood serum from diphtheria-immunized livestock to diphtheria-infected livestock treated the latter.
The scientific community was already aware of substances in the body that prevent disease, thanks to the 1770s smallpox inoculation (vaccination) program. On the other hand, the ground-breaking publication above identified antibodies as the agents responsible for effective inoculation and demonstrated their promise as curative agents.
As a result, in addition to disease prevention, epidemiologists and other researchers may be able to use antibodies to cure ailments.
Other notable advances in antibody development include discovering the antibody structure and the identification of B-cells as the particular lymphocytes responsible for antibody production.
In addition, two researchers devised the clonal selection theory, which emphasizes how B-cells produce an antibody before releasing it to fight an antigen, contradicting the widespread belief at the time that the antigen serves as an antibody formation template.
However, the invention of the first bespoke monoclonal antibody in 1975 was a watershed moment in the history of antibodies.
By 2023, antibody production has shifted from hybridoma technology to more advanced phage display technologies. Furthermore, custom antibody services use techniques such as gene and peptide synthesis, protein expression, and antibody sequencing to suit the ever-increasing need in the monoclonal antibody custom service market.
Below is an overview of the steps in custom polyclonal antibody production.
Custom Polyclonal Antibody Production
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Peptide Design
Antibodies and antigens are natural proteins, and proteins are normally made up of amino acids and peptides. As a result, the initial step in peptide synthesis is to use antibody sequencing to generate a peptide that matches the properties of the target antigen. However, peptides have distinct characteristics, so the amino acid sequence must match the required qualities.
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Antigen Synthesis/ Purification
Following peptide design, the target antigen is generated from the synthesized peptides. Bespoke peptide synthesis companies are essentially one-stop shops that provide end-to-end services, including bespoke protein synthesis as part of the protein expression service. As a result, creating the target/custom antigen is simple.
However, in some circumstances, the entity seeking custom antibody synthesis services already has the target antibody. The peptide design and antigen synthesis stages are redundant in the case of an existing antigen. In such a circumstance, the first stage is antigen purification; an antigen must be at least 90% pure to function well in polyclonal antibody manufacturing.
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Inoculation
Custom polyclonal antibody synthesis, as opposed to custom monoclonal antibody production, is effective in more animal hosts than mice. Conversely, Rabbits are the preferred bespoke polyclonal antibody host because they are larger, allowing for more serum collection per host with fewer injections.
Second, the bunnies are easier to inject, making serum collection more compassionate. Chickens, conversely, emerge as favorable polyclonal antibody hosts because the antibodies develop in the eggs, allowing for non-invasive antibody retrieval.
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Serum Catching
After being exposed to the antigen, the antiserum is retrieved from the host animal. On the other hand, the time it takes to recover the serum is determined by how long it takes specific animal hosts to develop antibodies.
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Titer Analysis
A titer analysis determines the target custom polyclonal antibody amount in the collected antiserum. Titer tests can be administered using enzyme-linked immunosorbent assays (ELISA kits) developed by researchers.
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Antibody Purification
Purification of antibodies requires separating the desired bespoke polyclonal antibody from a crude organic mixture. To avoid denaturing the protein, the antibody production company employs a variety of procedures, including chromatography and mass spectrometry, depending on the target antibody’s features.
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Antibody Characterization
Characterization of custom polyclonal antibodies requires evaluating the purified antibody to determine its physical and chemical properties. It is a type of quality control and analysis to ensure that only the highest-quality custom antibodies make it into the order fulfillment batch.
The Benefits of Using Polyclonal Antibodies
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A Wide Range of Host Animals
Polyclonal antibodies can be produced in various host species, such as rabbits, rats, chickens, goats, sheep, and horses. The quantity of antigen, injection methods and locations, adjuvant selection, and duration of immunization programs are all adjustable applications.
Because of the increased serum productivity and the animals’ longevity, huge animals with greater total blood volume will create enormous antibodies over time.
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High Stability
Polyclonal antibodies are a combination of antibodies with varying physicochemical properties, such as charge and hydrophobicity. Compared to monoclonal antibodies, they are more resistant to temperature and pH fluctuations. For the storage of monoclonal antibodies, stabilizing chemicals are usually necessary to prevent aggregation and precipitation and to sustain antibody binding.
Bottomline
As previously stated, polyclonal antibody manufacturing occurs by injecting antigens into specific lab animals such as goats, rabbits, and others. Even though these phases are essential to the overall approach, a researcher must comprehend the complete process, from animal immunization to antibody purification and quality control.
And after a few weeks, PAbs can be extracted from the antiserum. Making polyclonal antibodies is less expensive and more efficient than producing monoclonal antibodies.