Immunologic techniques polyclonal antibodies

This chapter deals with the application of immunological techniques to biochemical and molecular biological problems. We discuss the production of polyclonal antibodies and then a range of techniques that depend on the high specificity of antibody-antigen interactions. 

For triazine herbicides a very selective and sensitive immunological technique is recommended. The triazine herbicides are covalently bound to soil humic acids. A sandwich-immunoassay based on both polyclonal humic acid-antibodies and monoclonal triazine-antibodies111 was used for triazine detection. The schematic view of sandwich immunoassay is presented in Figure 4.3. The technique is very selective and sensitive. It can assure the best reliability of the analytical information because it does not require a prior separation. 

Production of antisera does not require specialized immunological techniques and cell culture facilities. Therefore it may be a simpler, faster and cheaper way than production of monoclonal antibodies (see section 3.3.). However, it must be noted that antisera-based immunoassays are sometimes less selective than monoclonal antibody-based immunoassay because of the polyclonality of antibodies in the antisera, and good antisera are not reproduced. The production of high affinity antisera may be helped by long-term immunization schedules from the mechanism of antigen-dependent B lymphocyte differentiation and selection (23). 

Immunological detection of ciguatoxins and related polyethers has received particular attention compared to other marine toxins. The initial RIA and enzyme immunoassay employing a polyclonal sheep anti-ciguatoxin antibody revealed cross-reactivities between ciguatoxins and other polyether toxins, suggesting the need for monoclonal antibodies. The availability of monoclonal antibodies allowed for the development of stick enzyme immrmoassay methods and solid-phase immunobead techniques (known as the paddle test), which successfully recognized toxins attached to correction fluid-coated bamboo sticks or paddles previously exposed to toxic fish tissues. 

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