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At least 127 records · Page 7Linked to original sources
PubMed1984

Assay for the specificity of monoclonal antibodies in crossed immunoelectrophoresis.

A method is described based on crossed immunoelectrophoresis of a complex antigen mixture in agarose gel followed by incubation of the gel with the monoclonal antibody. The bound monoclonal antibody is detected by the use of a secondary enzyme-labelled antibody. Using this technique we have been able to identify the precipitate arc in crossed immunoelectrophoresis of major histocompatibility complex (MHC) class I molecules in a mixture of all detergent solubilized cell membrane molecules by means of a monoclonal antibody, the specificity of which was known independently to be against MHC class I molecules. In other experiments using the same technique we demonstrated the reaction of a monoclonal antibody specific for chicken Ig light chains.

Animals↗
PubMed1986

Counter current line deflection immunoelectrophoresis. A technique for the quantification of antibodies.

Combining the electrophoretic principles of counter current immunoelectrophoresis and deflection of line precipitates in line immunoelectrophoresis provides a new technique for quantitative determination of antibodies against specific antigens (CCLD electrophoresis), even if no purified antigen or monospecific antibodies are available for construction of the detection system. We have used the method for quantitative determination of antibodies against the flagellum of Treponema phagedenis biotype Reiter and compared the diagnostic potential of this method in the diagnosis of syphilis with an ELISA method for the quantification of IgG antibodies against the flagellum. The CCLD electrophoresis could be optimized to a diagnostic performance very similar to that achieved using the ELISA method.

Animals↗
PubMed1980

Cerebrospinal fluid immunoelectrophoresis in multiple sclerosis.

In 95 patients with definite multiple sclerosis (MS) cerebrospinal fluid (CSF) cytomorphology, protein electrophoresis and immunoelectrophoresis using antisera to human serum, Fab fragments of immunoglobulin G(IgG) and to kappa and lambda light chains and the CSF IgG concentration were examined. In the CSF diagnosis of MS, the set of the examinations used is considered to be superior to any individual laboratory technique currently applied in the diagnostic process of MS. In the set of the CSF examinations, CSF immunoelectrophoresis revealed abnormal findings in patients with MS in whom presence of bands (oligoclonal gammopathy) in the gamma-globulin field of the CSF electropherogram and/or increased concentration of CSF IgG were not demonstrated.

Adult↗
PubMed1991

Neutrophil polarization and immunoelectrophoresis assays in the study of complement activation by biomaterials.

The neutrophil polarization assay, a technique used to measure chemotaxis, was adapted to examine complement activation. Complement activation in serum which had been incubated with metallic and polymeric biomaterials was examined using the neutrophil polarization assay and immunoelectrophoresis assay. In agreement with previous publications, nylon activated the complement cascade, but PTFE did not. The neutrophil polarization assay was found to be the most sensitive technique for examining complement activation by endotoxin but the immunoelectrophoresis assay is the technique most sensitive for detecting complement activation by cobalt powder. In both assays, complement activation was not detected in serum incubated with chromium powder. However, serum incubated with silver and nickel powder stimulated neutrophils to polarize indicating that these powders may activate complement.

Biocompatible Materials↗
PubMed1991

Heterogeneity of HDL in crossed immunoelectrophoresis with continuous gradient polyacrylamide gel in the first dimension and one or two antibody layers in the second dimension.

We describe a new method of immunoelectrophoresis with a continuous gradient polyacrylamide gel in the first dimension and an agarose-dextran gel in the second dimension with one or two layers of antibody. The use of a polyacrylamide gel in the first dimension allows better resolution of lipoproteins than with crossed immunoelectrophoresis using agarose gel in both dimensions. The use of two layers of antibody in the second dimension also enhances the specificity of characterization and the resolution of the separation. Thus, using a layer of anti-apo A-I combined with a layer of anti-apo A-II, three particles containing only apo A-I and three containing both apo A-I and A-II could be separated.

Adult↗
PubMed1989

Three different types of alpha-fetoprotein in the diagnosis of malignant solid tumors: use of a sensitive lectin-affinity immunoelectrophoresis.

A sensitive new technique for lectin-affinity immunoelectrophoresis was applied to samples from 28 infants and children in order to distinguish the origin of elevated alpha-fetoprotein (AFP) in sera. This new immunoelectrophoresis was successfully performed within 24 hours in sera with AFP as small as 910 ng/mL. With combined use of concanavalin A (Con A) and lentil agglutinin (LCH) binding tests, AFPs were classified into three subtypes: benign hepatic condition type (six patients), hepatocellular carcinoma type (nine patients) and yolk sac type (12 patients). AFP was of hepatocellular carcinoma type in all seven patients with hepatoblastoma, and of benign hepatic condition type in six of seven patients with elevated AFP due to conditions such as hepatitis, biliary atresia, and normal newborn. The question as to whether AFP produced in "hepatoblastoma" is of benign hepatic condition type or hepatocellular carcinoma type was first answered by the information in this present report. The differentiation between yolk sac and general hepatic AFPs was completed with the Con A binding test.

Adolescent↗
PubMed1976

Detection of plasmacytoma of the nasal cavity by immunoelectrophoresis of nasal washing fluid.

A patient with a solitary IgG-lambda plasmacytoma of the nasal cavity is described. Immunoelectrophoresis of the nasal washing fluid showed the presence of the same paraprotein. After radiotherapy the tumour and the paraprotein disappeared. Recurrence of the tumour after 6 months was again associated with the presence of the paraprotein in the nasal washing fluid. Immunoelectrophoresis of nasal washings may offer an easy method in detection and follow-up of localized plasmacytomas of the upper respiratory tract.

Aged↗
PubMed1975

Subtilopeptidase A isoenzyme system. Interaction with serum components and its importance for quantitative immunoelectrophoresis.

A method was developed which involved electroimmunoassay and crossed immunoelectrophoresis of subtilopeptidase A (EC 3.4.21.14). Initial trials with unfractionated antiserum were not successful and interaction of the enzyme with non-immunoglobulin serum components were shown to be the cause of the failures. Quantitative immunoelectrophoresis was possible when purified immunoglobulins were used. A pH of 6.5 (lower than the usual pH 8.6) was necessary to obtain a proper baseline definition. Subtilopeptidase A was confirmed as a multiple isoenzyme system. Qualitative inter-batch variations were detected. Di-isopropyl phosphorofluoridate inhibition altered the electrophoretic pattern, but no loss of antigenic determinants was observed.

Animals↗
PubMed1981

Immunochemical measurement of serum prostatic acid phosphatase (PAP). Clinical evaluation of radioimmunoassay and counter immunoelectrophoresis.

Two radioimmunoassay procedures (RIA-1 and RIA-2) were evaluated for the quantitation of prostatic acid phosphatase in serum and compared with the enzymatic method and counter immunoelectrophoresis method for their specificity and sensitivity. Sera from 168 patients were analyzed and these included: normals, 27; untreated prostatic cancer patients Stage A, 2; Stage C, 3; Stage D, 17; cancer of prostate treated with different modalities, 42; sarcoma of prostate, 1; prostatitis, 3; nonprostatic carcinoma, 17; and benign prostatic hyperplasia (BPH), 56. RIA-1 procedure appeared more sensitive (82% sensitivity) and specific (94.5% specificity) than the RIA-2 procedure (68% sensitivity and 91.8% specificity), but the differences were not statistically significant. The enzymatic method was found to be least sensitive (63.6% sensitivity) but also the most specific (100% specificity). Only 69 of the specimens were analyzed by counter immunoelectrophoresis, which showed sensitivity of 87% and specificity of 51.4%. False positives were observed more often in patients with nonprostatic cancer and BPH. The variations in diagnostic specificity of immunologic assays suggest the need of characterization of each antibody specificity.

Acid Phosphatase↗
PubMed1981

Rocket immunoelectrophoresis of C4 and C4d. A simple sensitive method for detecting complement activation in plasma.

Activation of complement component C4 has recently been measured by the quantitation of C4 and C4d (a cleavage fragment of C4) by electroimmunodiffusion in gels containing specific precipitating antibodies for C4 and C4d (Rocket immunoelectrophoresis, RIE). Quantitative measurements of the complement component C4 and its fragment C4d were determined in rocket immunoelectrophoresis (RIE) and compared with measurements of total hemolytic complement activity (CH50) or concentrations of C4 as determined by single radial immunodiffusion (RID). This newly developed RIE assay shows activation of the classical complement pathway and involves electroimmunodiffusion in gels containing specific precipitating antibodies for C4 and its cleavage fragment, C4d. In 37 plasma samples, excellent correlation was demonstrated between the C4 in RIE and CH50 (r = .70) and C4 by RID (r = .87). In vivo activation of C4 was determined by measuring the ratio of C4d to C4; 21 of the plasma samples assayed had ratios greater than 1.1 indicating activation of C4. In 13 of the plasma samples there were correspondingly low CH50 values, whereas 8 had normal CH50 levels. Therefore, activation can be detected in those instances when other measurements (CH50 and C4 quantitation) are normal. Thus the RIE assay for plasma C4 activation appears to be the most sensitive method available for assessing in vivo activation of the classical pathway of complement.

Animals↗
PubMed1978

Crossed immunoelectrophoresis of a herpes simplex virus type 1-specific antigen: immunological and biochemical characterization.

Crossed immunoelectrophoresis of herpes simplex virus (HSV) antigens obtained from infected cells solubilized with the nonionic detergent Triton X-100 has shown that antigen number 6 (Ag-6) is found in preparations from HSV type 1-infected cells but not in HSV type 2 preparations. A monospecific antiserum was produced against Ag-6 by intracutaneous inoculation of the corresponding immunoprecipitate into rabbits. This antiserum neutralized HSV type 1 but not HSV type 2. The polypeptide composition of Ag-6 (molecular weight, 131,000 daltons) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitate, using radiolabeled antigen. The multitude of information obtained demonstrates that crossed immunoelectrophoresis can be used preparatively, since specific immunoprecipitates represent a short cut for the attainment of immunoreactive viral protein entities useful for further immunological and biochemical analyses.

Antigens, Viral↗
PubMed1978

Quantification of influenza virus structural proteins using rocket immunoelectrophoresis.

The major influenza A virus structural antigens, matrix protein, nucleoprotein, haemagglutinin and neuraminidase were measured rapidly and accurately using a rocket immunoelectrophoresis technique. Virus was disrupted with 1% (w/v) sodium sarcosyl and electrophoresed into agarose containing specific antiserum to the individual virus structural proteins in 0.05 M-barbitone buffer, pH 8.6, for 1 to 4 h. For haemagglutinin antigen assays statistical analysis indicated that the coefficient of variation within an immunoelectrophoresis plate was 8.0% for antigen concentrations in the range 15 to 40 microgram/ml protein. For haemagglutinin and matrix protein the method was sufficiently sensitive to measure concentrations of antigens as low as 1.5 and 2.0 microgram/ml respectively. By incorporation in the agarose of mixtures of antisera against specific antigens of the virus, haemagglutinin, matrix or nucleoprotein could be assayed simultaneously.

Antigens, Viral↗
PubMed1984

Crossed immunoelectrophoresis and enzyme-linked immunosorbent assay of the cell-surface antigens of Bacteroides fragilis.

Antisera were raised to whole, live cells of a reference strain (NCTC 9344) and two clinical isolates (GNAB 92 and GNAB 4) of Bacteroides fragilis. Each antiserum was reacted in crossed line immunoelectrophoresis (CLIE) with EDTA-heat-sonication-prepared outer membrane (OM) complex from 10 B. fragilis strains. In addition, the antisera were reacted with these antigens in an enzyme-linked immunosorbent assay (ELISA). In CLIE, the antisera raised to the reference strain and one of the clinical isolates (GNAB 92) demonstrated a heat labile antigen which was common to all 10 of the test strains. Lipopolysaccharide (LPS) prepared from both the clinical isolates produced three major precipitin lines when reacted with their homologous antisera in crossed immunoelectrophoresis (CIE). In both cases, these three antigens were present as major components of the OM complex. Each antiserum reacted significantly in ELISA with all test OM complex preparations. Inhibition of ELISA showed that carbohydrates were the predominant cross-reactive antigens in ELISA and that in the case of the clinical isolate GNAB 4, most of the cross-reactive antigenic activity was present in the homologous LPS preparation.

Antigens, Bacterial↗
PubMed1983

Use of Aspergillus fumigatus mycelial antigens in enzyme-linked immunosorbent assay and counter-immunoelectrophoresis.

Antigens obtained from the ruptured mycelium of Aspergillus fumigatus were separated on the basis of their attachment or non-attachment to Concanavalin A-linked Sepharose gel. They were analysed by double-diffusion and two-dimensional immunoelectrophoresis with rabbit antiserum and sera from patients with aspergillus-related disease. Their sensitivity was assessed in an enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G. Sera which gave very low or high extinction readings on ELISA gave, in general, comparable results on counter-immunoelectrophoresis; but weak electrophoretic reactions were not closely paralleled by ELISA results. The unfractionated extract and the fraction bound by Concanavalin A-Sepharose had similar ability to detect specific IgG in sera from patients with aspergillosis.

Antigens, Fungal↗
PubMed1985

Allergy to guinea pigs: II Identification of specific allergens in guinea pig dust by crossed radio-immunoelectrophoresis and investigation of the possible origin.

An extract of dust from the air-vent filters of a room housing guinea pigs was analysed by quantitative immunoelectrophoretic procedures and compared with extracts of various materials derived from guinea pigs. Crossed radio-immunoelectrophoresis (CRIE) of the dust, performed with sera from twenty asthmatic patients who were positive by skin testing and RAST to guinea pig extracts, identified fourteen IgE-binding constituents. Although responses varied, most sera reacted with four particular allergens, antigens 2, 3, 10 and Sl. The numbers of allergens recognized by individual patients correlated with the RAST score, but not with total serum IgE. All seventeen dust constituents detected by crossed immunoelectrophoresis (and all four major allergens), were also present in extracts of guinea pig dander, fur, saliva and urine; several of these components were absent in an epithelial extract, and there were even less in preparations of shaved pelt, serum or faeces. None of the dust extract antigens were detected in materials used in animal husbandry, dust samples from rooms without guinea pigs, or a D. pteronyssinus extract. These findings suggest that inhalant allergens may be derived predominantly from material shed from the guinea pig coat after contamination with saliva, and possibly to a lesser extent, urine.

Allergens↗
PubMed1989

The allergens of dog. I. Identification using crossed radio-immunoelectrophoresis.

The antigens present in an extract of dog hair and dander were examined by crossed immunoelectrophoresis (CIE) and the IgE-binding allergens by crossed radio-immunoelectrophoresis (CRIE), respectively, using sera from 60 British and Finnish animal-allergic subjects. The extract was comprised of a minimum of 28 antigens, 11 of which were common to dog serum. IgE antibody in the sera of the dog-sensitive patients bound to 21 of the 28 antigens at varying frequencies and intensities. Binding of any intensity occurred most frequently to two serum proteins: antigen 23 (IgG) binding IgE in 88% of cases, and antigen 3 (dog serum albumin, DSA) in 77% of cases. Dander antigen 8 bound in 63% and antigen 1 in 42% of the sera. Strong IgE binding, however, was most commonly associated with dander antigen 8 followed by antigens 1 and 23 (IgG) then 3 (DSA). The ranking of the antigens as allergens was similar for the two populations except that DSA was more important for the British than for the Finnish subjects.

Allergens↗
PubMed1983

Analysis of Leptospira antigens by crossed immunoelectrophoresis.

Serovars icterohaemorrhagiae, canicola, and arboreae of pathogenic Leptospira interrogans and serovar patoc of saprophytic L. biflexa were examined by crossed immunoelectrophoresis. A close antigenic relationship was found between the interrogans serovars, particularly between icterohaemorrhagiae and canicola. To a much lesser extent cross-reactions were found between interrogans serovars and patoc. Comparison of three different antigenic preparations of icterohaemorrhagiae by tandem crossed immunoelectrophoresis and by absorption experiments with the patoc reference system showed the presence of several common broadly reactive antigens. Antigen 1 of the reference system, a genus-specific heat-stable protein antigen, was found in all antigenic preparations. It is a diagnostically important antigen.

Animals↗