Polyclonal Peptide Antisera

Abstract

Polyclonal antibodies are relatively easy to produce and may supplement monoclonal antibodies for some applications or even have some advantages. The choice of species for production of (peptide) antisera is based on practical considerations, including availability of immunogen (vaccine) and animals. Two major factors govern the production of antisera: the nature of adaptive immune responses, which take place over days/weeks and ethical guidelines for animal welfare. Here, simple procedures for immunization of mice, rabbits, sheep, goats, pigs, horses, and chickens are presented.

Peptides, Antibodies, Peptide Antibodies and More

“…As such, PAbs either represent naturally occurring Abs or Abs induced upon immunization. Basically, PAbs are produced by injecting an immunogen into an animal along with an adjuvant [87]. Adjuvants are used in the immunization to enhance the immune response [88,89].…”

“…Aluminumhydroxide, Al(OH) 3 , is often favoured since it has relatively few side effects and effectively activates the innate immune system, especially the alternative complement pathway, which leads to opsonisation and activation of the adaptive immune system and, ultimately, to production of Abs with high titres [88,89]. The most common host animal for production of PAbs is the New Zealand white rabbit, but larger animals (e.g., goat, sheep, and horse) are often used for large scale production [87]. Subcutaneous injections are preferred, since they are effective and least harmful to the animal.…”

“…Subcutaneous injections are preferred, since they are effective and least harmful to the animal. After being injected with a specific immunogen, the animal is usually given further immunizations (boosting) to ensure Abs of high titers against the specific target [87]. After immunization and boosting, PAbs are obtained, while recognizing different epitopes of an antigen.…”

“…Köhler and Milstein pioneered the technique for producing monoclonal Abs (MAbs) of desired specificities [92,93]. The initial steps for generation of PAbs and MAbs are similar [86,87,94]. The immunogen is injected into the host along with an adjuvant, which is followed by repetitive immunizations to boost the immune response of the immunized animal.…”

“…Amino acid composition Hydrophilic aas, charged aas, Pro, Gly (represented in loops) [33,138] Peptide length 8-25 aa [46,130,[132][133][134] Peptide structure Linear, flexible, cyclic, loops, turns, helices [25,86,87,[93][94][95] Protein target Accessible epitope, areas of high conservation, areas of hypervariability, N/C-termini, post-translational modifications [128,129,138] Following peptide synthesis, the peptides are conjugated to carriers to ensure immune responses with high antibody titres, since many peptides are not immunogenic by themselves. Traditional carriers include bovine serum albumin, keyhole limpet hemocyanin, and ovalbumin [45,128,[139][140][141].…”