Select monoclonal antibody or polyclonal antibody, the first thing is to determine the use of the antibody you need.
Polyclonal antibody is derived from multiple B cell clones, can be widely used in: Western blot; Radioimmunoassay (RIA); Enzyme-linked immunosorbent assay(ELISA); Indirect/direct immunofluorescent antibody test; Erythrocyte agglutination test; Immunohistochemistry(IHC); Immunology precipitation(IP); Immune diffusion and Affinity chromatography, etc.
Besides, polyclonal antibody’s production time is shorter and cost is lower than monoclonal antibody’s.
Main points during production
The types of antigen include peptide, recombinant protein, cell, plasmid DNA, etc. The quality and type of antigen you selected are directly related to whether you can get the specific and practical antibody you need.
The following points should be considered in selecting the most suitable animal model: the genetic relationship between antibody and antigen; use of the antibody; specificity of antibody reaction; the quantity of serum you need; the age and sex of animal.
To the production of polyclonal antibody, adjuvant can be divided into antigen delivery system (include emulsion and liposome) and immunopotentiator (like muramyl dipeptide). By now, there are more than 100 kind of adjuvants and the most widely used adjuvants include: Freund’s adjuvant, TiterMax, RAS, Gerbu, Imject.
The ultimate use of antibodies determines the purification method. If the purity requirement is not high, precipitation method is rapid and inexpensive. If the antibody is used for diagnosis, protein A/G purification is suitable. And if the antibody requires high-purity and high-specificity, antigen affinity purification can be used.
The main detection methods include: SDS-PAGE, western blot, isoelectric focusing and ELISA test. Except the final product needs to be detected, the titer of serum antibody also needs to be detected during production.