Anti-albumin Antibodies Research Articles

Homogeneous fluoroimmunoassay using Lucifer yellow VS: determination of albumin plasma.

Non-separation fluoroimmunoassays are well suited to automation. The potential sensitivity of such assays has not been realised because the most commonly used label (fluorescein) has absorption and emission spectra which coincide with those of blood constituents and because it has a very small Stokes shift. Lucifer yellow VS has a larger Stokes shift than fluorescein and an emission maximum at longer wavelength. We describe an energy transfer fluoroimmunoassay for plasma albumin in which Lucifer yellow VS is used to label albumin and rhodamine B isothiocyanate is used to label anti-albumin antibodies. The assay shows good correlation with a dye-binding method for albumin and has sensitivity and precision which compare favourably with similar assays using a fluorescein label.

Antibodies from patients with liver diseases and from normal human or animal sera against glutaraldehyde-polymerized albumins: lack of species specificity.

By the use of glutaraldehyde-polymerized albumins of different species (human, rabbit, bovine, goat and mouse) it was demonstrated that anti-albumin antibodies in sera of patients with liver diseases and in normal human and animal (rabbit, mouse) sera are not species specific.

Immunochemical characterization of anti-albumin antibodies in liver diseases

Anti-albumin antibodies (AAA) were isolated from sera of hepatic patients and normal individuals by affinity chromatography on insolubilized glutaraldehyde-treated human albumin. Anti-albumin antibodies were found to belong to IgG and IgM classes in both normal and hepatic patients. The normal level of AAA increased in pathologic conditions, the increase recorded for IgM AAA being higher than that for IgG AAA. The dissociation rate of AAA from the radiolabeled antigen in normal and hepatic sera showed that the affinity of AAA was higher in normal sera than in sera of patients with chronic liver disease and acute viral hepatitis. Anti-albumin antibodies were fractionated into two populations (AAA 1 and AAA 2) by a two-step chromatographic procedure. AAA 1 and AAA 2 were found different as regards their affinity for the antigen; specifically, AAA 1 affinity was higher than that of AAA 2. The other difference between AAA 1 and AAA 2 might stand in their specificity for the haptenic and structural determinants present in the glutaraldehyde-treated albumin.

Virus-Associated Receptors for Polymerized Human Serum Albumin in Acute and in Chronic Hepatitis B Virus Infection

The receptor for polymerized human albumin, recently identified on the hepatitis B virus, was studied by hemagglutination in 89 patients with acute and chronic hepatitis B virus infection. The receptor, which was differentiated from antialbumin antibodies by immunofluorescence and hemagglutination inhibition, was detected mainly in patients with active virus replication, independently of hepatitis B surface antigen titer. Thirteen of 20 uncomplicated acute hepatitis B cases were positive at clinical onset (mean log2 titer +/- SD: 4.20 +/- 2.46) but became negative during the acute phase, while 5 patients who progressed to chronic hepatitis had significantly higher receptor titers at onset (7.0 +/- 0.0, p less than 0.01) and remained positive afterwards. In chronic hepatitis B virus infection, the receptor was detected in 17 of 30 patients with chronic active hepatitis, in 5 of 12 with chronic persistent hepatitis, and in 2 of 16 healthy carriers. Among hepatitis B e antigen positive cases a wide range in receptor levels was observed, and in longitudinal studies most patients with low initial titers seroconverted to hepatitis B e antibody while patients with higher titers remained persistently hepatitis B e antigen positive. These results indicate that the virus receptor for polyalbumin may represent a useful marker in hepatitis B e antigen positive acute and chronic hepatitis B.

Anti-albumin antibodies in normal rabbit sera

Antibodies reacting with homologous glutaraldehyde (GA)-modified albumin were demonstrated in normal rabbit sera (NRS) by passive hemagglutination and direct binding assay. The ability of rabbit anti-albumin antibodies (AAA) to react with homologous GA-modified albumin was found to increase with the degree of albumin polymerization. AAA did not react with GA-treated monomeric albumin. It was proved that rabbit AAA are not species-specific since they react also with heterologous GA-polymerized albumin. The possible role of AAA in the catabolism of in vivo aged albumin molecules, antigenically similar with GA-induced albumin polymers, is discussed.

C1q has albumin binding activity but does not interfere in the hemagglutination test for anti-albumin antibodies

Two factors in the sera of patients with liver disease interact with polymerized human serum albumin (HSAP). These are anti-albumin antibodies (AAA) and receptors for polymerized albumin on HBsAg. Recently it was demonstrated by radioimmunoassay that purified human C1q also binds HSAP. Our data confirm the reaction between purified human C1q and HSAP by passive hemagglutination and by indirect immunofluorescence of HSAP-coated erythrocytes (E-HSAP). It is shown that although purified C1q binds HSAP, in the serum, the AAA and not C1q are responsible for the albumin binding activity in HBsAg-negative sera of patients with liver disease and of normal individuals, as detected by passive hemagglutination of E-HSAP. AAA were found to inhibit the binding of serum C1q to polymerized albumin, and hence the E-HSAP hemagglutination test for AAA titration in human sera proved valid.

Evidence for the separation of albumin- and apo A-I-dependent mechanisms of cholesterol efflux from cultured fibroblasts into human plasma.

The role of albumin has been studied in the plasma-mediated efflux of cholesterol from cultured fibroblasts. Immunoaffinity chromatography of plasma on immobilized anti-albumin antibody decreased by 25-50% total efflux catalyzed by plasma. The remainder of the efflux-promoting capacity of plasma was deleted by immunoaffinity chromatography on antibody to apolipoprotein A-I, the major apoprotein of high density lipoprotein. Both components of efflux were saturable with half-saturation at 0.5-1.0% (v/v) plasma. However, the net transport of sterol from cells to medium catalyzed by lecithin:cholesterol acyltransferase, was not reduced by the deletion of the albumin-catalyzed component of efflux. This finding was confirmed with congenitally analbuminemic plasma. These results indicate that efflux to albumin and to high density lipoprotein in plasma represent independent mechanisms; only the latter is coupled to net transport.

Antibodies against glutaraldehyde-modified albumin in normal mouse sera

An antigen-coated plate radioimmunoassay was used to detect antibodies against homologous glutaraldehyde-modified albumin (in monomeric or polymeric form) in normal mouse sera. Mouse antibodies reacted also with heterologous glutaraldehyde-treated albumin; for instance human albumin. Immunoelectrophoresis of purified anti-albumin antibodies and adsorption experiments on protein A-Sepharose 4B gel indicated that mouse antibodies belong mainly to the IgG class. The ability of mouse anti-albumin antibodies to react also with pyridinium compounds suggested that such structures are probably part of the new antigenic determinants induced in mouse albumin by glutaraldehyde treatment. It was assumed that anti-albumin antibodies have a physiologic role in the recognition and removal from the circulation of in vivo ‘aged’ mouse albumin.

Anti-albumin antibodies in sera of patients with gastro-intestinal disease.

Sera from 111 patients with various gastro-intestinal (GI) diseases were studies for the presence of antibodies to human serum albumin (HSA), bovine serum albumin (BSA) and ovalbumin (OA) by passive haemagglutination assay. The antibody titre to BSA was higher than that to HSA or OA. The anti-BSA antibody was demonstrated in upper GI diseases i.e. esophageal cancer, gastric ulcer, gastric cancer and duodenal ulcer, and not in lower GI disease i.e. Crohn's disease, ulcerative colitis and colon cancer. Both the mean titre and the incidence of the anti-BSA antibody tended to be higher in women than in men, and the titre was in a positive correlation with serum gamma-globulin levels. Sephadex G-200 column chromatography revealed that the anti-BSA antibody was widely distributed between void volume and 7S fraction.

Immune recognition of serum albumin. 15: Localization by Synthesis of antigenic Site 4 of bovine serum albumin to the ` region around the disulfide bond 166–175

Recently, we have localized and confirmed by synthesis the regions within which reside five major antigenic sites of bovine serum albumin and propoed that the remaining sixth major antigenic site (antigenic site 4) was localized within subdomain 3 (fragment 115–184) of albumin. In the present work, antigenic site 4 was localized to fall around the disulfide bond 166–175 by synthesis and immunochemical reactivity of the region 162–179. The synthetic 18-residue peptide was shown to bind substantial amounts of antibodies from early (38-day) and late (398-day) anti-albumin antisera from two rabbits. As much as half the total anti-albumin antibodies could be bound by the peptide from the late antisera. It was concluded that antigenic site 4 resides within, but may not include all of, the region 162–179 of albumin.

Shared idiotypes among anti-albumin antibodies of different species.

A system designed to detect plasma cells that produce antibodies directed at autologous idiotypic determinants of anti-human serum albumin (HSA) antibodies in rabbits was used to determine whether anti-HSA antibodies of horse, goat, swine and chicken origin were cross-reactive with rabbit antibodies of the same specificity. Fluorochrome-tagged anti-HSA preparations of these diverse species were used to stain splenic plasma cells of HSA-immunized rabbits and a similarly immunized chicken. The degree of idiotypic cross-reactivity, as detected by binding of anti-HSA antibodies to anti-idiotype within plasma cells of HSA-immunized animals, was sometimes equal to autologous staining. However chicken anti-HSA, the most phylogenetically distant idiotype examined, was demonstrably less cross-reactive than that obtained from the other species. Likewise, chicken plasma cells usually did not bind mammalian anti-HSA antibodies to an appreciable degree, as compared with autologous staining. These findings provide evidence for serologic and possibly structural similarities of antibodies of the same specificity from different species.

HBsAg-associated albumin receptors and antialbumin antibodies in sera of patients with liver disease

Several investigators reported that sera from patients with liver disease react with polymerized human serum albumin. This reactivity may be mediated by antibodies to polymerized albumin or polymerized albumin receptor sites on HBsAg. We tested sera from 114 patients with a variety of liver diseases by techniques which differentiate polymerized albumin binding activity mediated by HBsAg-associated receptors from that due to immunoglobulins. Total polymerized albumin binding activity was detected by passive hemagglutination of polymerized albumin-coated red blood cells, in 62.3% of patients with liver disease. The specificity for polymerized albumin was proven by hemagglutination inhibition of 18 sera using polymeric, monomeric, and native albumin as inhibiting antigens. Indirect immunofluorescence of polymerized albumin coated red blood cells after incubation with serum revealed that antibodies, HBsAg or both mediated polymerized albumin binding. The predominant immunoglobulin class with polymerized albumin binding activity was IgG in sera from patients with chronic active hepatitis and alcoholic liver disease, and IgM in acute viral hepatitis, chronic persistent hepatitis, and primary biliary cirrhosis. HBsAg-mediated polymerized albumin binding activity was frequently associated with antibodies to polymerized albumin. The attachment of hepatitis B virus to hepatocytes may be facilitated by polymerized albumin via polymerized albumin receptors. Antibodies to polymerized albumin may interfere with this binding and may result in reduced infectivity.

Serum antibodies to albumins in normal human pregnancy.

era from 72 pregnant women were studied for the presence of antibodies to human serum albumin (HSA), bovine serum albumin (BSA) and ovalbumin (OA) by passive haemagglutination assay. Anti-albumin antibodies were found in 96% of the cases and distributed throughout the 3 trimesters of pregnancy.

Immunological approach to the study of neurohumoral sleep factors: Effects on REM sleep of antibodies to brain stem proteins

Proteins were collected for several months from the reticular formation of some 40 cats. The sodium dodecyl sulfate electrophoretic profile showed eight visible bands. Rabbits were inoculated with these proteins four times at weekly intervals in order to produce antibodies. Antibodies were also produced against cat serum proteins and cat serum albumin. These antibodies were tested for their effects on the sleep-wake cycle of another group of cats. The results of these experiments showed that the anti-reticular formation (MRF) antibodies and not antiserum or antialbumin antibodies, produced a specific decrease in rapid eye movement (REM) sleep without altering slow-wave sleep. It was also observed that when the anti-MRF antibodies were incubated with MRF proteins, the effect was lost, but that it persisted when they were incubated with serum protein only. It was also noted that the reduction in REM sleep was the result of an increase in the latency and a decrease in the frequency of REM periods, rather than in the duration of individual periods. In addition, the antibodies decreased some of the phasic elements of REM sleep such as multiple-unit and eye movement bursts. These experiments present the first direct evidence suggesting that specific protein molecules play an important role in triggering REM sleep.

The effect of binding and eluting conditions on the immunoaffinity purification of rabbit anti-albumin antibodies.

Conference Article| August 01 1980 The effect of binding and eluting conditions on the immunoaffinity purification of rabbit anti-albumin antibodies M. ISLA HALLIDAY; M. ISLA HALLIDAY 1Department of Biochemistry, The Queen's University, Belfast BT9 7BL, Northern Ireland, U.K. Search for other works by this author on: This Site PubMed Google Scholar G. BRIAN WISDOM G. BRIAN WISDOM 1Department of Biochemistry, The Queen's University, Belfast BT9 7BL, Northern Ireland, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1980) 8 (4): 430–431. https://doi.org/10.1042/bst0080430 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation M. ISLA HALLIDAY, G. BRIAN WISDOM; The effect of binding and eluting conditions on the immunoaffinity purification of rabbit anti-albumin antibodies. Biochem Soc Trans 1 August 1980; 8 (4): 430–431. doi: https://doi.org/10.1042/bst0080430 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search Keywords: IgG, immunoglobulin G This content is only available as a PDF. © 1980 Biochemical Society1980 Article PDF first page preview Close Modal You do not currently have access to this content.

Antibodies against nickel-albumin in rabbits and man.

Five albino rabbits were immunized every 2 weeks by intradermal injections of a nickel rabbit-albumin complex and during alternate weeks by occluded cutaneous applications of 5% nickel sulphate in petrolatum. After 6 weeks of immunization, passive immunodiffusion disclosed precipitating antibodies against the nickel-albumin complex in the serum of the rabbits. The specificity of these antibodies was substantiated by the precipitation of anti-albumin antibodies with rabbit albumin prior to immunodiffusion. Passive immunodiffusion was used to examine serum from 14 nickel-sensitive patients for precipitating antibodies against nickel human-albumin complex. Five of the patients had widespread erythematous, non-eczematous reactions following oral challenge with 2.5 mg nickel as nickel sulphate, while five others suffered an aggravation of their nickel dermatitis. Precipitating antibodies were found in three patients, all of whom had experienced rapid-onset erythema after oral challenge. A weak precipitate was seen in one of nine controls. The specificity of the precipitates was substantiated by reproducibility and by negative immunodiffusion after precipitation in the positive sera with the antigen.

Deeper insight into HBsAg--anti-albumin antibody correlations.

The presence and titers of anti-albumin antibodies (AAA)--ie, precipitins (AA-IP) and agglutinins (AA-Aggl)--and the serum concentrations of immunoglobulins and albumin were determined in 210 asymptomatic carriers of HBsAg grouped according to HBsAg titer and subtype. A different immunologic pattern was observed in the HBsAg/ad and HBsAg/ay carriers, the results suggesting an increased aggressivity for the HBsAg/ay subtype which was evident in subjects with lower HBs antigenemia and characterized by higher concentrations of IgG and IgM and rises in AA-P titers. A positive linear correlation was found between HBsAg and AA-Aggl titers; the carriage of HBsAg/ad was associated with significant higher values of AA-Aggl than of HBsAg/ay. These subtle correlations between HBsAg titer and subtype and AAA suggest that the human serum albumin (HSA) known to exist on the HBsAg particles may be similar in a modified form, to that representing AAA specificity.

Interaction of hepatitis B surface antigen with polymerized human serum albumin

HBsAg binds to a solid-phase adsorbent consisting of polymerized human serum albumin (HSA) on glass particles. Both AD and AY antigenic subtypes of hepatitis B surface antigen (HBsAg) display this interaction. In either case, the binding to polymerized HSA is reduced in the presence of human serum, suggesting significant attachment of serum components at the locations on HBsAg particles where polymerized HSA binds. The temperature dependence of the interaction goes through a maximum above room temperature, in contrast to the increasing reaction with temperature of the HBsAg--anti-HBs antibody system. The interaction between HBsAg and polymerized HSA is discussed in relation to previous findings of HSA polymers and anti-polymerized albumin antibodies in hepatic patients. A mechanism for production of an autoimmune, antialbumin antibody condition, in association with hepatitis B virus infection is proposed.

Isolation and characterization of nascent albuminyl peptides synthesized in vivo from rat liver

A procedure is described for the isolation of nascent albuminyl peptides from rat liver polysomes which is based on the isolation of total peptidyl tRNA by ion-exchange chromatography on ECTEOLA cellulose followed by immuno-affinity chromatography employing monospecific anti-albumin antibodies immobilized on Sepharose 4B. Identity of the isolated nascent albuminyl peptides was assayed by tryptic peptide fingerprint analysis. Quantitation and determination of the specific activity of the nascent albuminyl peptides, labeled in vivo with l-[ 14C]leucine, were made by subjecting the peptides to acid hydrolysis, dansylation and resolution of the amino acids by thin-layer chromatography, and determination of the specific activity of dansyl leucine.

Autoantibodies detected in rabbits hyperimmunized with group A, C, and G streptococcal vaccines

Rabbits hyperimmunized with group A, C, and G streptococcal vaccines developed autoantibodies with affinities for different tissues (smooth muscle, cytoplasmic, and myocardial antibodies) and for autologous proteins (anti-albumin and anti-immunoglobulin antibodies). The presence of anti-albumin and smooth muscle antibodies, associated with a high level of immunoglobulin, suggests the development of hepatic disorders in the hyperimmunized rabbits.