Sam M. Beiser

Antigenicity of Steroid-Protein Conjugates

Testosterone, cortisone, deoxycorticosterone, estrone, and progesterone act as haptens when they are conjugated with bovine serum albumin. Antibodies with steroid specificity are formed in rabbits immunized with each of the five steroid hormone-protein conjugates.

Antibodies to small molecules: biological and clinical applications.

Publisher Summary This chapter discusses the methods of eliciting antibodies, detection of antibodies, determination of specificity of antibodies, and iminunoassay methods. The development of sensitive methods for detecting and quantitating reactions between hapten and antibody and the perfection of techniques for obtaining highly purified preparations of antihapten antibodies have played a significant role in the development of modern immunological concepts. Antihapten antibodies have proved particularly useful in the study of antigenic determinants, structural requirements for immunogenicity, the nature of antigen-antibody reactions, and the chemical, physical, and biological properties of antibody. In recent years, the haptens of biological importance have been studied with increasing frequency, and the chapter discusses the production, detection, characterization, biological properties, and clinical applications of antibodies with specificity for such haptens.

Immunochemical Detection of Minor Bases in Nucleic Acids

Rabbits immunized with bovine serum albumin conjugates of 5-bromouracil, 5-iodouracil, and 6-methyladenosine produced antibodies specific for the bases. These antibodies were used to detect immunochemically 5-bromouracil and 6-methyladenosine in denatured DNA.

Immunochemical studies on a galactosyl-protein conjugate

Antibodies formed in rabbits immunized with galactosido- β - D -azophenyl bovine serum albumin, after absorption with bovine serum albumin, were studied by quantitative hapten-inhibition techniques. It was shown (a) that the surface profile of the antibody is specific for the β - D -galactopyranosyl structure, (b) that the dimension of the combining site is complementary to the galactopyranosyl-phenylazo moiety and is approximately half that of the previously well-defined human antidextran site, (c) that the antibody or spectrum of antibodies in sera from two rabbits present an apparent micro-identity.

Hypersensitivity to purified trypsin and chymotrypsin.

A NUMBER of proteolytic enzymes have gained wide acceptance for clinical use.1 They are employed in the treatment of peripheral vascular disease, particularly superficial thrombophlebitis, for the relief of bronchial obstruction due to inspissated secretions in patients with chronic pulmonary disease, for the debridement of body surfaces covered with necrotic tissue, for the reduction of hematomas and edema resulting from trauma, in various acute ophthalmic inflammatory conditions and as an adjunct to cataract surgery.2 Commercial preparations include trypsin or chymotrypsin or both, derived from mammalian pancreas, incorporated in sesame oil for intramuscular injection, in tablets for oral administration, in nose .xa0.xa0.

Comparison of antibodies and enzymes specific for β-d-glucosides

Abstract Rabbit antisera to β- d -glucosylphenylazo-bovine serum albumin were obtained and were absorbed with bovine serum albumin to remove to remove the anti-protein antibodies. The specificity of the antibody combining region was determined by hapten-inhibition experiments. The dimensions of this combining site encompass the glucosylphenylazoaryl moiety of the antigen. A comparison of the antibody-combining ability of aryl- and alkyl-glucosides with the activities of the same compounds as substrates, inducers, or complexers for microbial β- d -glucosidases, revealed that the effect of changes in orientation of the sugar hydroxyls and of the glycosidic link is roughly the same; that alkyl-glucosides are bound less strongly than aryl-glucosides in glucosyl-specific proteins; that the strong inhibitory ortho effect of substituents on aromatic aglycones for binding by microbial glucosidase is exhibited by the antibody-antigen system; and that electronic effects by various substituents on aryl glucosides are comparable for the antibody and for emulsin. These results provide additional evidence for homology between the combining regions of antibodies and enzymes which are induced by the same determinant groups.

Immunochemical detection of 7-methylguanine residues in nucleic acids

Abstract The ability of specific antibodies to react with 7-methylguanine residues in nucleic acids was investigated. Anti-7-methylguanine specific antibodies precipitated polymers of poly-guanylic acid which were methylated to an extent of 35 or 70% at the N-7 position of guanine, indicating that these antibodies could readily detect 7-methylguanine residues in a polynucleotide. This reaction was proportional to the total amount of 7-methylguanine present, suggesting further that quantitation of these residues is possible. To determine the minimal amount required for detection, varying amounts of 7-methylguanine were introduced into calf thymus DNA by alkylation with dimethyl sulfate. While showing no reaction with denatured nonalkylated DNA, the reaction of antibodies with alkylated DNA was proportional to the amount of 7-methylguanine in the preparations. Moreover, the antibodies appeared to detect differences in the distribution of 7-methylguanine residues in extensively methylated DNA. Precipitation was observed with DNA containing as little as one 7-methylguanine residue per 300 nucleotides, suggesting that these antibodies can be used to detect biologically significant levels of 7-methylguanine in viral and cellular nucleic acids.

Antibodies specific for purine and pyrimidine nucleosides: Effect on in vitro priming ability of DNA☆

Antibodies specific for thymine and guanine nucleosides react immunochemically with denatured but not with native DNA. These antibodies inhibit the ability of calf thymus DNA to serve as a primer for DNA polymerase extracted from chick embryos. No inhibition of DNA synthesis is observed with anti-bovine serum albumin or normal sheep globulin. Anti-T is more inhibitory than anti-G whether Bacillus cereus, Micrococcus lysodeikticus or calf thymus DNA preparations are used as primers. Purified anti-T inhibits the priming of DNA to the same extent, relative to antibody content, as the globulin fraction anti-T. The data are consistent with the hypothesis that binding of the anti-nucleoside antibodies to the exposed bases in denatured DNA prevents these masked regions from acting as templates for the DNA polymerase.

Purification of Antibody to Galactosyl-Protein Conjugates

A rapid procedure for purification of rabbit antibody to β-D-galacto-sylphenylazo-bovine serum albumin is described. Antibody was precipitated with a heterologous antigen and was then dissociated from the precipitated complex with hapten. The antigen was partially removed by pH adjustment, and antibody was separated from residual antigen and from hapten by partition chromatography on Sephadex G-25. The antibody, obtained in 50 to 55 percent over-all yield, was 96 percent reactive with homologous antigen. It appeared to consist, of 95 percent of a single component, based upon physicochemical measurements.

Studies of materials derived from staphylococcus: III. Protective action of gamma globulin against the anaphylactoid effects of a staphylococcal bacterial fraction

Abstract Through use of the agar precipitin method, human sera have been shown to contain antibodies directed against both culture filtrates and bacterial extracts of staphylococci. Most of the antibodies were directed against culture filtrates. All of the Staphylococcus aureus strains studied produced antigens which reacted with HGG, while only two of nine Staphylococcus albus strains formed antigens reacting with HGG. This suggests a relationship between the pathogenicity of a strain of staphylococcus and its ability to stimulate antibody production.