The general procedure to produce polyclonal antibodies is as follows: An antigen/adjuvant conjugate is injected into an animal of choice to initiate an amplified immune response.
After a series of injections over a specific length of time, the animal is expected to have created antibodies against the conjugate.
Animals frequently used for polyclonal antibody production include chickens, goats, guinea pigs, hamsters, horses, mice, rats, and sheep.
Animal selection should be based upon: Goats or horses are generally used when large quantities of antisera are required.
Chicken IgY does not fix mammalian complement component C1 and it does not perform as a precipitating antibody using standard solutions.
When using rabbits, young adult animals (2.5–3.0 kg or 5.5–6.5 lb) should be used for primary immunization because of the vigorous antibody response.
Female rabbits are generally preferred because they are more docile and are reported to mount a more vigorous immune response than males.
The size, extent of aggregation and relative nativity of protein antigens can all dramatically affect the quality and quantity of antibody produced.
Injection of soluble, non-aggregated proteins has a higher probability of inducing tolerance rather than a satisfactory antibody response.
In general, microgram to milligram quantities of protein in adjuvant are necessary to elicit high titer antibodies.
The affinity of serum antibodies increases with time (months) after injection of antigen-adjuvant mixtures and as antigen in the system decreases.
Antigens should be free of preparative byproducts and chemicals such as polyacrylamide gel, SDS, urea, endotoxin, particulate matter and extremes of pH.
Expasy has aggregated a set of public tools under its ProtScale page that require some degree of user knowledge to navigate.
This includes some of the more common characteristics: Investigators should also consider the status of nativity of protein antigens when used as immunogens and reaction with antibodies produced.
Selection of specific adjuvants or types varies depending upon whether they are to be used for research and antibody production or in vaccine development.
[1][2][3] Rho(D) immune globulin is made from pooled human plasma provided by Rh-negative donors with antibodies to the D antigen.
PAbs remain stable in different environments, such as a change in pH or salt concentration, which allows them to be more applicable in certain procedures.
Additionally, depending on the amount needed, PAbs can be made in large quantities in relation to the size of the animal used.
Despite the drawbacks, polyclonal antibodies' broad epitope recognition[10] and higher sensitivity[11] are key across different applications.
[12] Recent technology advancements allowed the direct sequencing of polyclonal antibody proteins from human and immunized animals.
[13][14] Those antibody proteins can then be recombinantly produced to mimic the original pAbs generated by the immune system.