PubMed Health⌕ Search

Biomedical subjects

L Miribel

Publications and source records attributed to L Miribel.

14 recordsLinked to original sources
PubMed1988

Electrotransfer of proteins following polyacrylamide gel electrophoresis. Nitrocellulose versus nylon membranes.

We found that some plasma proteins such as haptoglobin (Mr 200,000) and alpha 1-acid glycoprotein (Mr 40,000) do not bind at all to nitrocellulose membranes in classical transfer conditions. In contrast, these proteins can be clearly identified when a charged nylon membrane is disposed on the anodal side of the nitrocellulose membrane. Several factors which are important in modifying the adherence between proteins and the blotting membrane have been studied. They are not directly related to the molecular weight of the protein considered but also include the presence of organic solvents in the buffer systems, the pore size of the membrane and the degree of reduction and/or glycosylation of the protein. The eventuality of decreased or absent identification of some antigens on nitrocellulose membranes stresses the importance of controls using nylon membranes.

Blood Proteins↗
PubMed1988

Ceruloplasmin.

Explore the source record for details and available documents.

Ceruloplasmin↗
PubMed1988

The use of dye-ligand affinity chromatography for the purification of non-enzymatic human plasma proteins.

Literature data are analysed in this review on the use of immobilized triazine dyes for the characterization, isolation and purification of non-enzymatic human plasma proteins in both conventional and high-pressure liquid chromatography systems. Attention is focused on the mode of interaction between the dyes and these proteins, as well as on the advantages over previously reported techniques. Future developments are discussed.

Blood Proteins↗
PubMed1987

Purification of human alpha 1-beta glycoprotein and study of its microheterogeneity and molecular variants.

Alpha 1-beta glycoprotein (A1B) was purified to homogeneity from human plasma using a three-step procedure involving pseudo-ligand affinity chromatography on immobilized Cibacron Blue 3-GA, gel filtration chromatography on Sephadex G-200 and ion-exchange chromatography on DEAE Affigel Blue. The overall yield of the combined techniques was 31%. The major advantages of this technique include its conveniency, and the fact that no apparent alteration of the purified protein occurred as judged by the absence of modification of its physicochemical parameters. A1B appears to be present in normal plasma as a major form of Mr 70,000 daltons with extensive charge heterogeneity, together with minor components of higher Mr.

Blood Proteins↗
PubMed1987

Fractionation of plasma proteins by pseudo-ligand affinity chromatography. Elution from immobilized Brown Fractogel TSK-AF.

The fractionation of human plasma by chromatography on immobilized Brown Fractogel TSK-AF was analyzed by following the elution profile of 25 different plasma proteins. A three-step procedure was used to elute proteins from the column. First, a low-molarity buffer (30 mM sodium phosphate, pH 7.0, I = 0.053) was applied; then a linear salt gradient (0-1.0 M sodium chloride in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M sodium chloride. Tightly bound proteins were finally stripped with 0.5 M ammonium thiocyanate. The elution profile of the proteins obtained by this procedure appears to be very reproducible. Comparison with the profiles obtained by chromatography on Cibacron Blue 3-GA and on Green TSK-AF indicates significant differences between the binding properties of the three gels.

Animals↗
PubMed1987

Fractionation of plasma proteins using dye-ligand chromatography: elution profiles on immobilized green TSK-AF.

The fractionation of human plasma by chromatography on immobilized Green TSK-AF was assessed by immunological analysis of the elution profiles of 27 different plasma proteins. A three-step procedure was used to elute proteins from the column. First a low-molarity buffer (30 mM sodium phosphate, pH 7.0, I = 0.053) was applied; then a linear salt gradient (0-1.0 M NaCl in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M NaCl. Tightly bound proteins were finally stripped with 0.5 M NH4SCN. The elution profile of the proteins using this procedure appears to be very reproducible. Comparison with the profile obtained upon chromatography on Cibacron Blue 3GA [Gianazza, E. and Arnaud, P. (1982) Biochem. J. 201, 129-136] indicates significant differences between the binding properties of the two gels. These differences can be used to design a "tandem-chromatography" system which provides an efficient means for the separation of several plasma proteins.

Blood Proteins↗
PubMed1987

Alpha 1-acid glycoprotein (alpha 1-AGP) on the membrane of human lymphocytes: possible involvement in cellular activation.

A glycoprotein termed alpha 1-acid glycoprotein (alpha 1-AGP) is a component of normal human serum; its concentration is often increased in several pathological disorders, including acute inflammation and cancer. Inhibitory effects of alpha 1-AGP on some in vitro T and B cell function assays have been reported but our recent data indicated that alpha 1-AGP is indeed a T cell mitogen at physiological concentrations. The present study was designed to investigate: (a) the relationship between this glycoprotein and two other glycoproteins of the T and B cell membrane, i.e. the T3 and Ia antigens; (b) the ability of lymphocytes to take up exogenous alpha 1-AGP; (c) the different expression of alpha 1-AGP on the T cell membrane upon different activation pathways, i.e., autologous non-T-cells (B cells and monocytes) phytohemagglutinin and anti-T3 monoclonal antibody (MAb) stimulations. The data reported herein show no competition at the membrane level between anti-alpha 1-AGP and anti-T3 or anti-Ia MAbs. In addition, (1) the lymphocytes were able to absorb alpha 1-AGP from the culture medium and (2) the expression of this glycoprotein was enhanced upon T cell stimulation (all three stimulants employed induced an increase of alpha 1-AGP positive T cells), thus suggesting a possible role of this glycoprotein in in vitro T cell activation.

Adult↗
PubMed1987

Purification of IgM monoclonal antibody from murine ascitic fluid by a two-step column chromatography procedure.

Two IgM monoclonal antibodies (Mabs) were purified from murine ascitic fluid by a two-step procedure involving pseudoligand affinity chromatography on immobilized Cibacron Blue F3GA and gel filtration chromatography on ACA-22. The purified IgM from one ascites sample, H-1, was determined to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis while ascites containing E3.1 IgM Mab, was purified to greater than 95% IgM. The retention of biological activity was confirmed by immunofluorescence staining of peripheral blood lymphocytes.

Acrylamides↗
PubMed1986

Effect of isoprinosine on sialylation of interleukin-2.

The effects of Isoprinosine (ISO) on interleukin-2 (IL-2) production by human peripheral blood mononuclear cells (PBMC) were investigated. Treatment (of human PBMC) with ISO enhanced IL-2 production by PBMC from 7 of 10 normal individuals. However, no augmentation of IL-2 production was observed when cultures of HUT-78 cells, a human leukemic T cell line, were treated with ISO. IL-2 purified from supernatants of human PBMC treated with ISO exhibited pI values of 5.5 and 6.4. IL-2 prepared from untreated PBMC exhibited a single pI value of 8.2. The pI value of IL-2 prepared from ISO-treated PBMC shifted to 8.2 after treatment with neuraminidase, demonstrating that the IL-2 molecules isolated from ISO-treated PBMC possessed sialic acid. The pI values of the IL-2 isolated from ISO-treated and untreated HUT-78 culture supernatants were identical (pI = 7.8) and were not modified by neuraminidase treatment. These results suggest that the increase in IL-2 production following treatment of PBMC with ISO may be mediated through the activation of a distinct subset of IL-2 producing cells. Furthermore, the sialylation of IL-2 may be of physiologic and immunopharmacologic importance.

Adjuvants, Immunologic↗
PubMed1984

A three-step purification of human alpha 1-acid glycoprotein.

alpha 1-Acid glycoprotein (AGP) was purified to homogeneity by a 3-step procedure using pseudo-ligand affinity chromatography on immobilized Cibacron blue F3GA, Procion red HE3B, and preparative column isoelectric focusing. The overall yield of the combined techniques was 88%. Analysis of the purified AGP by lectin affinity chromatography on immobilized Con A and immunoaffino-electrophoresis indicated that the most acidic form did not interact with the lectin, while the two more basic fractions possessed different affinities for Con A. In addition, 3 different populations of AGP were clearly separated by Con A affinity chromatography.

Chromatography, Affinity↗