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At least 19 recordsLinked to original sources
PubMed1978

[Localization of mitochondrial cytochrome b using specific antibodies].

Specific antibody has been obtained against cytochrome b (pig heart mitochondria). It inhibits the electron transport of the respiratory chain in the intact mitochondria at the cytochrome b site of the inner mitochondrial membrane. It has no effect on the isolated submitochondrial particles which are inside-out inner membrane vescicles free of any outer membrane or outside-out inner membrane. These findings indicate a probably not transmembranous topologic localization of cytochrome b; this component of the respiratory chain seems located near the outer side of the inner mitochondrial membrane.

Animals↗
PubMed1976

Natural infection of man with group A streptococci. Levels; restriction in class, subclass, and type; and clonal appearance of polysaccharide-group-specific antibodies.

The level of antibodies specific for the two serologically distinct polysaccharides of group A and group A-variant streptococci was investigated in human sera by a modified Farr technique. Whereas there was an obvious relationship between the anti-streptolysin O titer and the level of group-A-specific antibodies, suggesting a recent streptococcal infection with group A streptococci, this correlation was not apparent for the level of group A-variant antibodies. There is evidence suggesting that children have significantly lower group A polysaccharide antibody levels than adults; this is not true of their group A-variant antibody levels. There was an apparent restriction of groups A- and A-variant-specific antibodies to the kappa light-chain type and IgG2 heavy-chain subclass. Whereas only one serum contained group A-specific antibodies also associated with IgG3 heavy chains, nine sera contained anti-group A-variant-specific antibodies with both IgG2 and IgG3 heavy chains. No anti-polysaccharide activity was detected in the IgM and IgA class of serum antibodies. Anti-group A and anti-group A-variant antibodies of human sera are restricted with respect to the number of clonotypes that are shown by isoelectric focusing. The most heterogeneous sera contained roughly ten clonotypes.

Antibodies, Bacterial↗
PubMedDate not supplied

Analysis of anti-sperm sera for T/t locus-specific antibody.

An objective antiglobulin radioimmunoassay for the analysis of anti-spermatozoa antisera has been used to characterize more than 30 alloantisera. Each of these antisera was raised against spermatozoa from mice carrying T/t regin mutations. Whilst all of the antisera had considerable activity for mouse spermatozoa, none of them was found to contain antibodies specific for T/t region gene products. A detailed analysis of a single serum revealed that its activity is for antigens which are non-polymorphic, sperm-specific and species-specific. The failure in this study to detect antibodies specific for T/t region gene products is discussed.

Absorption↗
PubMed1979

Divergent results in radial immunodiffusion. III. Isolation and specificity test of class-specific antibodies.

A method is described for the isolation of antibodies specific for immunoglobulin class determinants from antisera containing antibodies both for the immunoglobulin class and for its subclasses, using affinity chromatography. The model system used is bovine IgG with its subclasses IgG1 and IgG2, together with antisera raised in goats and sheep. The specificity of the isolated class-specific antibodies was demonstrated by double diffusion (DI) and radial immunodiffusion (RID). The levels of total IgG in normal bovine sera measured with the class-specific antibodies were the same as the sum of the IgG1 and IgG2 levels obtained with subclass-specific antisera. However, sera from cattle infected with Fasciola hepatica contain IgG1 fragments without class-specific determinants. In these sera, contrary to expectation, the class-specific antibodies gave higher total IgG levels than the subclass-specific antisera. The yield of class-specific antibodies varied with different batches of antisera. The minimum amount of each class-specific antibody preparation needed for a RID plate also varied. Class-specific antibodies isolated from antisera raised against IgG1 gave more intense precipitation reactions in RID against IgG1 than IgG2 and vice versa. Possible reasons for these differences are discussed.

Animals↗
PubMed1976

Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated chicken muscle myosins.

Antibodies prepared against actomyosins can be shown to behave similarly, if not identically to more recently prepared antibodies against highly purified myosins. Details of the purification of the antigens, and of the production of antibodies to chick myosins from smooth gizzard muscle and from striated pectoral muscle are given. The antibody specificity appears to be directed against the heavy chains of the myosin molecules, since these antibodies specifically inhibit the myosin ATPase reaction, and since in situ staining of myosin polypeptide chains on an SDS gel using the antibodies in indirect fluorescence shows staining only in the heavy band region. Use of the antibodies in immunofluorescence microscopy suggest that the antibodies are tissue, but not species, specific. Example of their use in staining tissue sections are shown.

Actomyosin↗
PubMed1975

Topological studies on the hydrolases bound to the intestinal brush border membrane. II. Interactions of free and bound aminopeptidase with a specific antibody.

The position of the intestinal brush border aminopeptidase with respect to the lipid bilayer has been investigated with the aid of right side out vesicles prepared from the brush border and an immunological technique using an unlabelled or peroxidase-labelled antibody specific for aminopeptidase. The finding that the bound form of the enzyme was almost as readily inhibited and agglutinated as the free form during incubation with the antibody was consistent with the view that the majority of the aminopeptidase surface emerged from the bilayer. This finding was entirely corroborated by the observation that only a few antigenic determinants were not free to react with the antibody in bound aminopeptidase. This immunological technique may be applied to other membrane proteins provided that preparations of the pure proteins and of specific antibodies are available.

Aminopeptidases↗
PubMed1977

Specific antibody levels in the intestinal perfusates of Heligosomoides polygyrus-infected mice.

Radial immunodiffusion analysis was performed on intestinal perfusates from ICR/CD1 mice following oral infection and challenge with viable Heligosomoides polygyrus larvae. Infection of these mice resulted in an alteration in the amount and class of immunoglobulins, as well as anti-parasitic antibody. Specific IgA peaked at 3 days post-infection and then showed a gradual decline until day 21. This rapid elevation in IgA levels was temporally associated with early events in the parasite life cycle, and the increase in IgA was significantly higher in mice infected twice with H. polygyrus prior to challenge. Differences in both the concentrations and kinetics of specific IgA and IgG1 were also observed in the perfusates from these mice when compared to those noted for animals infected only once.

Animals↗
PubMed1978

[Mitochondrial localization of cytochrome c1 with specific antibodies].

A specific antibody against cytochrome c1 (pig heart mitochondria) has been obtained. It inhibits the electron transport of the respiratory chain in the intact mitochondria at the cytochrome c1 site of inner mitochondrial membrane ; but it has no effect on the isolated submitochondrial particles (inside-out inner mitochondrial membrane vesicles free of any outer membrane or outside-out inner membrane). Thus the topologic position of cytochrome c1 in the inner mitochondrial membrane is asymetrically lcoated on the outer side of the inner mitochondrial membrane. These results agree with our previous researches on ATP-ase and cytochromes b, c and a, indicating the location on the inner side for the first one, transmembranous for the last one, on the outer side for the others respiratory chain components. Thus the electron transport from cytochrome b to a takes place in the outer region of inner mitochondrial membrane and the transmembranous location of cytochrome-oxidase facilitates the transfer of the electrons to oxygen.

Animals↗
PubMed1977

Larva-specific antibodies in patients with visceral larva migrans.

Seven of 10 patients with visceral larva migrans (VLM) had serum precipitating antibodies specific for larval antigens of Toxocara canis as determined by double diffusion in agar. Such antibodies were absent in 114 normal adults and 25 patients with collagen disease. Precipitation of ascarid components by C-reactive protein resulted in false-positive reactions, but this precipitation was readily prevented by appropriate absorption of sera before testing. A more sensitive assay, the enzyme-linked immunosorbent assay, revealed high titers of antibodies to larvae of Toxocara in all patients with VLM; the log2 titer ranged from 9 to 14. Five of eight household relatives of these patients and four children who had had VLM two to four years before testing had titers of 6-12. Of the 114 normal adults, 105 had titers of 0-2; nine had titers of 3-8. Of the 25 patients with collagen disease, 23 had titers of 0, and two had titers of 4 and 6, respectively. Additional studies of those patients indicated that infection with Toxocara can be distinguished serologically from ascariasis, filariasis, and trichinellosis.

Antibodies↗
PubMed1979

A comparison of the ability of beta-galactosidase and horseradish peroxidase enzyme-antibody conjugates to detect specific antibodies.

The ability of two different enzyme-antibody conjugates to detect specific antibodies has been compared. beta-Galactosidase was conjugated to antibodies raised against rabbit Fc fragments using m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS). Horseradish peroxidase (HRP) was conjugated to part of the same batch of antibodies using the periodate method. The beta-galactosidase and HRP labels enabled detection of approximately 8 fmoles and 4 fmoles respectively of human growth hormone (HGH) antibodies, when their enzyme activities were measured spectrophotometrically. The detection limit of the beta-galactosidase label was increased 4-fold when a fluorimetric detection system was employed.

Animals↗
PubMed1975

The antibody-enzyme analogy. Comparison of enzymes and antibodies specific for phosphopyridoxyltyrosine.

Reduced Schiff base compounds of pyridoxal-P and tyrosine, which were used to induce specific antibodies described in the preceding article (V. Raso and B. D. Stolar, Biochemistry, 1975), caused active site-directed inhibition of tyrosine transaminase and tyrosine decarboxylase. The antibodies, studied as analogs of enzymes, were able to bind an unsaturated Schiff base catalytic intermediate, as shown by equilibrium dialysis and absorbance difference spectroscopy. Schiff base formation can proceed while the pyridoxal-P and tyrosine are within the antibody combining site, but the rate of this bimolecular condensation within the sites was not greater than the rate in free solution. Antibody did effect a small rate enhancement for the pyridoxal-P-catalyzed transamination of L-tyrosine. These results are discussed in light of current ideas in the mechanisms of enzyme catalysis.

Animals↗
PubMed1979

The demonstration of antibody specificity by a new technique. The gel electrophoresis-derived enzyme-linked immunosorbent assay (GEDELISA) and its application to antibodies specific for feline leukemia virus.

By combining the high resolution of sodium dodecylsulfate polyacrylamide gel electrophoresis with the sensitivity of enzyme-linked immunosorbent assay (ELISA) antibodies specific for different feline leukemia virus components are characterized. Based on the same principle, Concanavalin A binding sites of FeLV components are also detected.

Antibodies, Viral↗
PubMed1977

Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin.

The injection of rabbits with insoluble or soluble G-actin from chicken smooth or striated muscle will produce antibodies that are equally reactive, and species and tissue non-specific in immunoprecipitation, immunofluorescence and actin-activated Mg2+-ATPase inhibition tests. These antibodies have been used for the identification of actin-containing fibrils in a variety of tissues. When G-actins from chicken smooth or striated muscle are immobilized by chemical linkage to Affi-Gel 702 microbeads, their immunogenicity is increased, but the antibodies obtained against them are species-specific and will only react with actin and actin-containing structures from chicken and are therefore limited in use. It is concluded from this work that insoluble G-actin is the preferable immunogen to obtain precipitating antibodies for wide use.

Actins↗
PubMed1979

Antigen-specific detection of soluble immune complexes by a solid phase specific antibody system.

An antigen-specific immune complex assay based on the following principle has been developed: xenogeneic, antigen-specific antibodies are attached to a solid phase. During first incubation with patient's serum, immune complexes in antigen excess are bound to the xenogeneic antibody by their free antigenic determinants. In a second incubation a labeled antibody, specific for human immunoglobulins, is combined with the antibody part of the immune complex. Quantitation of the label allows the determination of the immune complexes. The principle of the method has been varied using artificial soluble immune complexes of tetanus toxoid and human anti-tetanus toxoid antibody, and immune complexes prepared by mixing patient sera containing either HBsAg or anti-HBsAg antibodies. The reliability of the results is shown by their coefficient of variation (2.5%). With the method described soluble immune complexes predominantly in slight antigen excess, which are thought to be responsible for development of immune complex disease, have been detected.

Antibody Specificity↗
PubMed1979

Reversal of digitalis effects by specific antibodies.

Highly digoxin-specific or ouabain-specific antibodies can readily be obtained by immunizing rabbits or sheep with repeated injections of the glycoside coupled to protein carriers. By virtue of their binding capacity digoxin-specific antibodies are capable of removing digoxin concentrations from red blood cells and renal tissue specimens. As evidenced by various experiments with human erythrocytes and isolated cardiac preparations in vitro, digoxin effects are rapidly reversible in the presence of digoxin-specific antibodies. In vivo antidigoxin antibodies can protect animals from digoxin effects and promptly abolish established toxic effects, associated with marked alterations of digoxin pharmacokinetics. However, due to the large molecular weight, complete antibodies cannot be eliminated via the renal route. The use of antigen-binding (Fab) fragments of digoxin-specific antibodies offer the advantage of rapid renal elimination of bound and inactivated digoxin. So far, due to potential immune reactions, the clinical use of purified digoxin-specific antibodies of Fab fragments is restricted to life-threatening accidental or suicidal digoxin or digitoxin poisoning.

Animals↗
PubMed1976

Radioimmunoassay of oestrone in plasma. A comparison of different methods with respect to the relation between assay specificity, sample purification and antibody specificity.

A radioimmunoassay (RIA) for oestrone (Oe1) in plasma was developed using an ether extraction, a partition between NaOH and light petroleum, and a TLC as sample purification and an antiserum cross-reacting with Oe2 for the final assay (Method A1). The reliability criteria are given in detail. In order to simplify this method a highly specific antiserum was developed by using Oe1-3-hemisuccinate-BSA as an antigen. Using this antiserum for the final assay but omitting the TLC (Method B) the Oe1 concentration in male plasma was 76% overestimated (Method B compared with Method A1). In pregnancy plasma Oe1 could specifically be determined after a simple ether extraction (Method C). It was concluded that the use of a highly specific antiserum (as determined by cross-reaction studies) for the final assay does not necessarily imply that a chromatographic sample purification can be omitted without loss in assay specificity. This appears to be true mainly in cases where the steroid concentration of the sample is low. Normal values of Oe1 from 80 healthy adult males were ascertained by Method A1. Age group I (22-61 years, n = 48) ranged from 1.22-5.60 ng/100 ml, median 2.81; age group II (67-90 years, n = 32) from 1.55-6.40, 100 median 3.41. The small increase of Oe1 with age was highly significant.

Adult↗
PubMed1978

The prevention and reversal of digoxin intoxication with specific antibodies.

The formation of digoxin-specific antibodies was induced in sheep by immunization with a digoxin-albumin conjugate. The efficacy of the antibodies was investigated in anesthetized cats. When the digoxin-specific antibodies were administered prophylactically as a gammaglobulin, IgG or F (ab')2 preparation, the dose of digoxin needed to induce ventricular dysrhythmia was significantly greater (p less than 0.001) for the pretreated animals than for the controls. To investigate therapeutic efficacy, the animals were digitalized with digoxin over a period of three days and were given digoxin injections on the fourth day to provoke ventricular tachycardia. Of the control animals, three died before two hours had elapsed and the arrhythmia persisted in the two remaining animals. By contrast, a stable sinus rhythm was restored in all animals which were treated with F (ab')2 fragment of the digoxin-specific antibodies after onset of ventricular tachycardia. The doses of digoxin required to trigger renewed ventricular dysrhythmia in these animals were greater than those required at the start of the experiment. The potential clinical use of digoxin-specific antibodies is discussed in the light of these results and reports in the literature.

Animals↗