A Study of the Cross-Reactivity of Antipurin-6-Oyl Serum With Deoxyribonucleic Acid (Dna)

Abstract

The specificity of the reaction of antipurin-6-oyl sera with thermally denatured DNA has been studied by means of hapten inhibition techniques. The relative order of effectiveness of various haptens as inhibitors of the complement fixation reaction between DNA and antipurin-6-oyl serum was found to be comparable to their relative order of effectiveness as inhibitors of the precipitin reaction between purin-6-oyl-protein conjugates and antipurin-6-oyl serum. Antipurin-6-oyl antibody has been purified and has been shown to be capable of reacting with thermally denatured DNA. It is concluded that the reactivity of these purine-specific antisera with DNA is a property of antibody with specificity for the purin-6-oyl moiety. In addition, the reaction between antipurinoyl sera and DNA has been demonstrated by the techniques of radioimmunoelectrophoresis and passive cutaneous anaphylaxis.

Antigenicity of Polyribonucleotides*

“…However, the RNAase activity of the rabbit serum had to be neutralized by anti-RNAase before direct serological activity with RNA could be shown. Inhibition with RNA was noted by Stollar and Levine (1962) in a DNA-lupus antibody system and by Butler et al (1965) in the purin-6-oyl immune system. Antibodies to polyribonucleotides could prove extremely useful in probing the structure of RNA.…”

Immunological protection against digoxin toxicity

“…Control rabbits were immunized in a similar manner with BSA, purin-6-oyl-bovine serum albumin (Pur-BSA)conjugates (7) or with 0.85% saline solution alone in complete Freund's adjuvant mixture. The duration of immunization and the number of antigen injections are listed in Tables I and II 5%o HSA8 plus 93 ml Tris-buffered saline, pH 7.4 [10]) was added 64 mung digoxin-5H (randomly labeled; 126 g&c/ mg') in 2 ml of the same buffer. The reagents were mixed and incubated at 370C for 2 hr.…”

Biologic Activity of Digoxin-Specific Antisera

“…Purified y-globulin was also prepared in a similar manner from pooled normal rabbit Fraction II ' by dissolving it in 0.03 M phosphate, pH 7.0, and passing it over DEAE-cellulose. After concentration by negative pressure dialysis, each purified protein fraction was dialyzed against pH 7.45 Tris-buffered saline (21). Four of six preparations contained only proteins which migrated as -y-globulins on filter paper electrophoresis; the fractions from two anti-HSA-Dig sera (DB-38, DB-45) contained small amounts of ,6-globulin.…”