Polynucleotides, antibodies

With the exception of antibodies reacting only with native DNA all the different kinds of anti-polynucleotide antibodies described above can be induced experimentally. 

Murphy, R.E., Callans, S., Gonzales, M., Huyghe, B. (2005). Two-dimensional liquid chromatography for intact antibody analysis in serum. Presented at the International Symposium on the Separation of Proteins, Peptides and Polynucleotides. (ISPPP 2007) St. Pete Beach, FL. 

Fig. 1. Configuration of a microtiter plate used to study the induction of B-DNAtx> Z-DNA transition in poly(dG-m dC).poly(dG-m dC) by spermidine. The concentration of spermidine added to the polynucleotide (designated a to h and j inside the plate) is described in Table 1. Column 2 (DNA) contains poly(dG-m dC).poly(dG-m dC) similar to column 3. However, these wells will not be treated with the monoclonal antibody.

Subtract the mean background values from the mean values for wells conudning polynucleotide-spermidine complex. In the example given, subtract the mean obtained in Step 15 from that in Step 14. The result is the mean optical density owing to the binding of monoclonal anti-Z-DNA antibody to the polynucleotide. 

Procedures for preparation of the immunizing antigen, injection schedules, and assays for antibody formation are similar to those used for helical polynucleotide-MBSA antigens and are discussed in detail in the following section and in Vol. 12B [174] of this series. 

Antibodies specific for triple-helical polynucleotides clearly distin-... 

A primary dose of 50-200 fig of polynucleotide, in the MBSA complex and emulsified with complete adjuvant, is injected into each rabbit at several intradermal sites along the back and subcutaneously a total volume of 1 ml per rabbit is convenient. Similar doses, but with incomplete adjuvant, are given intradermally and subcutaneously on days 14 and 21, and the animals are bled 5-7 days later. They can be bled at weekly intervals, and additional booster immunizations can be given if antibody levels should fall. During early courses of immunization, they may peak and fall rapidly. ... 

There are three major obstacles that may limit the usefulness of polycarboxylates, polynucleotides and the tilorone congeners in clinical medicine toxicity, hyporeactivity and immunogenicity. The problem of immunogenicity has been most thoroughly investigated with ds-RNAs such as poly(I) poly(C). Injected parenterally into rabbits, poly(I) po y(C) and poly(A) poly(U) elicited the production of antibodies reactive with the ds—RNA 191 19S). 

Electrophoresis on a macro scale has been applied to a variety of difficult analytical separation problems inorganic anions and cations, amino acids, catecholamines, drugs, vitamins, carbohydrates, peptides, proteins, nucleic acids, nucleotides, polynucleotides, and numerous other species. A particular strength of electrophoi esis is its unique ability to separate charged macromolecules of interest to biochemists, biologists, and clinical chemists. For many years, electrophoresis has been the powerhouse method of separating proteins (enzymes, hormones, antibodies) and nucleic acids (DNA, RNA), for which it offers unparalleled resolution. ... 

In experimental studies on the formation of antibodies to a hapten-protein conjugate, it appears that the more immunogenic the protein, the better the response. Although proteins are usually the most efficient carriers, polypeptides, polysaccharides, lipid membranes, or even polynucleotides may be sufficient to impart im-munogenicity. In recent years, direct conjugation of simple chemicals to structural elements of cell membranes (especially of monocytes and macrophages) has been thought to play an important role in the induction of sensitization (de Weck 1975 Shearer 1975). The protein carrier or cell membrane structure of the host may contribute to the specificity of the response (partial autoimmunity). 

Plescia OJ, Braun W, Imperato S, Cora-Block E, Jaroskova L, Schimbor C (1968) Methylated serum albumin as a carrier for oligo- and polynucleotides. In Plescia OJ, Braun W (eds) Nuclei acids in immunology. Springer, New York Pressmann D, Goldberg AL (1968) The structural basis of antibody specificity. Benjamin, New York... 

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