Production of antisera against

The production of antisera against a cell type is one of the oldest effective methods of achieving a specific immune action. These antibodies can be divided into tw o groups polyclonal antisera that react with multiple antigenic determinants or epitopes and monoclonal antibodies that are directed at only a single epitope. 

A typical method for the production of antisera against digoxin is described below (22) ... 

In 1971 Chopra et al. (C8) published details on the production of AB against T4 (ranging in titer from 1 100 to 1 1000) produced in rabbits after injection of thyroglobulin. Others have produced antibody of much higher titer in rabbits (H13) and sheep (C20). These antisera were found to have satisfactory specificity for I-T4, although they do cross-react with d-T4, but this does not present any difficulty. 

Production and specificity of antisera against CF. CF -CF complex purified from Triton X-100 extract of thylakoids by anion-exchange chromatography and sucrose density centrifugation, and ATPsynthase complex from Triton X-100 dissolved thylakoids immunoprecipitated by anti CF showed the same subunit... 

The discovery of Lp(a) by Berg in 1962 (B6) relied on the production of rabbit antisera against beta-lipoprotein and on the selective absorption of these antisera with individual human sera. When certain human sera were used for absorption, the antisera retained precipitation capacity in radial immunodiffusion with 30-35% of individual human sera, which obviously contained a previous unknown antigen. The particle carrying the new antigen shared antigenic properties with beta-lipoprotein, but had an additional antigenic structure. This was evidenced from the only partial fusion of the precipitin bands formed between a positive human serum, the antibeta lipoprotein antiserum and the new absorbed antiserum. 

The production of IFN-y-neutralizing antibodies specific for an N-terminal peptide of human IFN-y provided the first evidence that the N-terminus of IFN-y contained an important functional site [27]. A similar approach was used to produce N-terminus-specific neutralizing antisera against murine IFN-y... 

Is nitrocellulose antigenic Some workers have been unable to achieve good results by immunization with nitrocellulose-bound protein. They reproducibly obtain antisera directed against nitrocellulose We found that in every case, this resulted from injecting powdered nitrocellulose m Freund s adjuvant using adjuvant actually increases the production of low affinity IgM that binds nonspecifically to nitrocellulose. We have never observed this effect in our experiments when the technique described here was followed strictly... 

Polyclonal antibodies are the serum product of an immunized animal containing many different antibodies against the various mixtures of antigens injected. The antiserum is the product of many responding clones of cells and is usually heterogeneous at all levels. These levels include the specificity of the antibodies, classes and subclasses, titer, and affinity. The response to individual epitopes may be clonally diverse, and antibodies of different affinities may compete for the same epitope. This variation means that polyclonal antisera cannot be reproduced see Fig. 3). 

Hekimi, S. and O Shea, M. (1989) Antisera against AKHs and AKH precursors for experimental studies of an insect neurosecretoiy system. Insect Biochem 19,79-83 Schneider, L. E., Sun, E. T., Garland, D. J, andTaghert, P H (1993) An immunocyto-chemical study of the FMRFamide neuropeptide gene products m Drosophila. J. Comp Neurol. 337,446-460... 

How can the differences in specificity of the antibodies obtained from rabbit, mouse and hamster be explained Although the antibodies obtained from animals of the last two species have not as yet been well characterized, they have in common the cross-reaction with poly A poly U. As the purine and pyrimidine bases involved in these complexes are different, they probably recognize the double-helical structure. It is quite unhkely that the polyribose phosphate chain plays an exclusive role in this specificity since we have seen that the anti-poly I poly C antibodies of rabbit react better with poly dG poly dC than with poly rG poly rC. We cannot exclude the possibihty that poly I poly C complexed to MBS A undergoes modifications when it is introduced into the bloodstream. The discovery by Stern in 1970 of an enzyme which specifically hydrolyzes double-stranded RNA or poly I poly C, and which exists at different levels in the sera of nine mammaUan species illustrates this possibihty. There will thus exist in the antisera antibodies against the products of degradation, the quantity varying according to the species. 

In 1895, Hericourt and Richet reported the first clinical trials testing the principle of antibody production. They injected cancer cells into animals to obtain antiserum to treat cancer patients this was the first time several patients with cancer were administered tailor-made serum for treatment. Several patients showed improvement, which was encouraging, but none of the patients were completely cured. These trials were repeated in the early 1900s but the results were not consistent. The problems included the variability of the antisera and the side effects of polyclonal antibodies - some of which were directed against self. 

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