Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0026764 (multiple myeloma)
36,148 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The polyethylene glycol-mediated cell fusion technique has been used to analyze the diversity of the antibody response to the terpolymer poly(Glu60Ala30Tyr10)(GAT). Nine stable clones (all producing IgM K anti-GAT antibodies) were isolated from a fusion between P3-X63-Ag8 myeloma cells and spleen cells from a DBA/2 mouse sensitized to GAT 5 days earlier. Seven other clones (producing IgG K anti-GAT antibodies) were derived from another fusion between NSI myeloma cells and spleen cells of (C57BL/6 X DBA/2)F1 hybrid mice hyperimmunized with GAT. These 16 anti-GAT antibodies were grouped according to their pattern of reactivity with GAT and the two related polymers of poly(Glu60Ala40) (GA) and poly(GLU50Tyr50) (GT). Two monoclonal anti-GAT antibodies (IgM F9-102.2 and IgG F17-148.3) demonstrated crossreactivity with GA but failed to crossreact with GT determinants. In contrast, the remaining 14 hybridoma antibodies demonstrated preferential reactivity with GAT but also exhibited crossreactive binding to GT and in some cases GA. There was a correlation between the fine specificity pattern and the presence of a common anti-GAT idiotype on these antibodies. Thus, the hybridoma anti-GAT antibodies which reacted with GT shared crossreactive idiotypic determinants (CGAT) present in mouse anti-GAT immune sera. In contrast, the monoclonal F9-102.2 and F17-148.3 antibodies that failed to bind to GT lacked the major CGAT idiotypic determinants.
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PMID:Fine specificity of antibodies to poly(Glu60Ala30Tyr10) produced by hybrid cell lines. 8 55

B leukemia cells from four different patients were hybridized with a mouse myeloma cell line with polyethylene glycol as a fusing agent. The original leukemia cells all expressed immunoglobulin on their surface, but failed to secrete it. Over 200 different human-mouse somatic cell hybrids were obtained; 57% of them secreted human immunoglobulin in large amounts. Human immunoglobulin secretion can be a stable property of these hybrid cells over months of continuous culture. In each case the human immunoglobulin secreted was restricted to the light chain type expressed by the parental B leukemia cell. In addition, these hybrid cells secreted the original mouse myeloma protein and a variety of mixed human-mouse immunoglobulin molecules.
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PMID:Rescue of immunoglobulin secretion from human neoplastic lymphoid cells by somatic cell hybridization. 9 69

The Fv fragment of mouse myeloma protein M313 was crystallized from poly(ethylene glycol) solution in the form of monoclinic crystals, space group C2 and unit cell dimensions a = 5.96 nm (59.6 A), b = 5.66 nm (56.6 A), c = 13.79 nm (13.9 A) and beta = 99.7 degrees. Some unusual effects of poly(ethylene glycol)on protein crystals were noted and are discussed.
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PMID:Crystallization of the Fv fragment of mouse myeloma protein M315. 49 96

Hybrids of BALB/c lymphocytes and a murine myeloma, a tumor that expresses intracisternal A-particles, were obtained with polyethylene glycol as the fusogen. The karyotype, tumorigenicity, and A-particle expression of the hybrid clones were assessed. All the hybrid clones analyzed were tumorigenic and expressed intracisternal A-particles even when they were the result of a fusion event between two lymphocytes and one myeloma cell in which no loss of chromosomes was detected. The tumors that developed following inoculation of hybrid cells into BALB/c mice (1 x 10(6) cells/mouse) were karyotypically identical to the inoculated cells. It appears that the two myeloma cell phenotypic traits analyzed (tumorigenicity and A-particle expression) are dominant.
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PMID:Tumorigenicity and intracisternal A-particle expression of hybrids between murine myeloma and lymphocytes. 49 79

A simple method is described for promoting the fusion of mouse myeloma cells in suspension with polyethylene glycol (PEG 1000). By carefully controlling the concentraion of PEG and the time of exposure of the cells, it was possible to obtain hybridization frequencies several-hundred-fold higher than those obtained with Sendai virus.
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PMID:A simple method for polyethylene glycol-promoted hybridization of mouse myeloma cells. 60 83

The chick kidney mitochondrial cytochrome P-450 1,25-dihydroxyvitamin D3 24-hydroxylase was partially purified by sequential polyethylene glycol precipitation, aminohexyl-Sepharose 4B, and hydroxylapatite chromatography. The specific activity of the final preparation, when reconstituted with NADPH, adrenodoxin, and adrenodoxin reductase, was 245 pmol/min/mg of protein or 0.56 pmol/min/pmol of P-450. The specific cytochrome P-450 content was 0.45-0.73 nmol/mg of protein. BALB/c mice immunized with this preparation developed serum polyclonal antibodies to the 24-hydroxylase, as demonstrated by immunoprecipitation. Splenic lymphocytes from an immunized mouse were fused with myeloma NSI/1-Ag-4-1 cells, and hybridomas secreting monoclonal antibodies to the 24-hydroxylase were detected by immunoprecipitation. The hybridoma lines were cloned by limiting dilution and further characterized as IgG1, IgG3, and IgM subclasses. In one-dimensional immunoblots of soluble 24-hydroxylase preparations, the monoclonal antibodies revealed a single band with an apparent molecular weight of 59,000. The monoclonal antibodies did not cross-react with cytochrome P-450s from other species but immunoprecipitated and immunoblotted a soluble chick renal mitochondrial 25-hydroxyvitamin D3 1 alpha-hydroxylase preparation, demonstrating the close similarity of these two hydroxylases. These antibodies were coupled to Sepharose CL-4B and used to isolate to homogeneity the two enzymes from chick kidney mitochondria. Amino-terminal sequences and amino acid composition data demonstrate that these enzymes are different but homologous.
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PMID:Immunopurified 25-hydroxyvitamin D 1 alpha-hydroxylase and 1,25-dihydroxyvitamin D 24-hydroxylase are closely related but distinct enzymes. 131 Jun 88

In this study we investigated whether electrofusion would enable efficient hybridization of human B cells to occur without the need of prior activation in vitro. Two cell lines, the murine myeloma SP 2/0 and murine-human heteromyeloma F3B6, were used as fusion partners, and hybridization of human B cells purified either from peripheral blood or from solid lymphoid tissue (tonsil) was studied. Optimal hybridization frequencies were obtained at a poration field strength of 3 to 4 kV/cm. Electrofusion appeared to be a very efficient method to hybridize tonsillar B lymphocytes, resulting on the average in a hydridoma outgrowth of 1 per 1400 lymphocytes in fusions with SP 2/0 and 1 per 3300 in fusions with F3B6. Hybridization frequency in polyethylene glycol-induced fusions was only 1 per 13,000 tonsillar lymphocytes. Seventy-five percent of the resulting hybridomas secreted human immunoglobulin, either IgM or IgG. Electric field-induced hybridization of peripheral blood B lymphocytes resulted in immortalization of 1 per 17,500 lymphocytes when using Sp 2/0 as a fusion partner, and 1 per 15,000 with F3B6 as a fusion partner. Polyethylene glycol fusions yielded only 1 hybridoma per 160,000 blood lymphocytes. On average, 60% of the hybridomas obtained by fusing peripheral blood B cells secreted human immunoglobulin. Both IgM and IgG secretors were found. Furthermore, by the use of a fusion chamber with a small volume (35 microliters), hybridomas could be generated from small numbers (300,000 to 55,000) of human B cells. In conclusion, human B cells can be hybridized by electrofusion with myeloma cells with efficiencies comparable to those found for murine splenocytes, without the need for prior activation in vitro.
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PMID:Efficient electric field-induced generation of hybridomas from human B lymphocytes without prior activation in vitro. 157 22

Peripheral blood mononuclear cells of an immunized patient were transformed with Epstein-Barr virus and then fused with P3X63Ag8 mouse myeloma cells by polyethylene glycol. After the cloning, a hybridoma cell line secreting specific anti-Jkb monoclonal antibody was isolated. The antibody was produced in supernatant form and tested for its use as a blood grouping reagent.
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PMID:A human anti-Jkb monoclonal antibody. 166 74

Two monoclonal IgG3 syngeneic anti-idiotypes are described which form soluble and insoluble complexes with anti-alpha(1----6)dextran hybridoma and myeloma proteins. Specific precipitation was seen when purified anti-alpha(1----6)dextrans were added to ascitic fluid containing IgG3 kappa anti-idiotype. Analysis of the supernatants of the idiotype-anti-idiotype precipitates demonstrated the presence of soluble complexes whose mobilities in polyacrylamide gels could, in some cases, be distinguished from that of free anti-idiotype. An IgG1 kappa anti-idiotype is described which did not form precipitates with anti-alpha(1----6)dextrans unless 3% PEG 6000 was added.
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PMID:An immunochemical analysis of precipitating and non-precipitating idiotype-anti-idiotype reactions. 169 51

Whilst monoclonal antibodies (Mab) to potyviruses have been generated, it has not been possible to produce molecules with high specificity or broad reactivity to defined conserved amino acid sequences. In the current study, peptide-mediated electrofusion was used to select for high efficiency antibody-secreting hybridomas after mice were immunized with highly immunogenic viral coat protein. Mice were immunized with coat protein from either one potyvirus (potato virus Y, PVY-D) or a mixture of five distinct potyviruses. Two well-defined peptides were used for selective electrofusions. Peptide-1 was selected from the highly specific N terminal region of PVY-D and peptide-2 from the highly conserved N terminal/core junction region of Johnson grass mosaic virus (JGMV). Conventional PEG-mediated fusions using mice immunized with these peptides did not result in hybridoma formation. On the other hand, electrofusions using biotin-streptavidin to bridge peptide-specific B cells to myeloma cells produced hybridomas secreting antibodies either highly specific to PVY-D or cross-reactive with all potyviruses, depending on the peptide used.
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PMID:The use of peptide-mediated electrofusion to select monoclonal antibodies directed against specific and homologous regions of the potyvirus coat protein. 171 98


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