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)

Three hybridomas producing monoclonal antibodies (McAb) against recombinant human granulocyte colony-stimulating factor (rhG-CSF) have been established by fusing mouse myeloma SP 2/0 cells with spleen cells from a BALB/c mouse immunized with rhG-CSF. Ascites was produced from BALB/c mice by injecting substrain 2D4-B4 hybridoma cells intraperitoneally. The antibody was purified either by the caprylic acid-ammonium sulfate method or by euglobulin precipitation. The caprylic acid-ammonium sulfate method was superior to euglobulin precipitation in terms of purification and recovery of IgG. The 2D4-B4 McAb was determined to belong to the IgG3 subclass with a molecular weight of about 172,400 It was proved by Western Blot to react specifically with rhG-CSF. The stability and affinity of the McAb were analyzed as well. The potential clinical and experimental applications of this McAb are discussed.
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PMID:[Studies on monoclonal antibody against recombinant human granulocyte colony-stimulating factor]. 171 45

Three hybridoma cell lines producing monoclonal antibodies (McAb) against recombinant human granulocyte colony-stimulating factor (rhG-CSF) have been established by fusing mouse myeloma cell line SP2/0 cells with spleen cells from a BALB/c mouse immunized with rhG-CSF. Ascites was obtained from BALB/c mice by injecting the hybridoma cells intraperitoneally. Three McAbs were purified by the caprylic acid-ammonium sulfate method. For each, the IgG subclass, valence and activity, molecular weight, specificity and affinity were determined. The applications of McAbs against rhG-CSF in the clinic and laboratory are discussed.
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PMID:Studies on monoclonal antibody against recombinant human granulocyte colony-stimulating factor. 172 68

The kidney mitochondrial monooxygenases known as 25-hydroxyvitamin D3 1 alpha- and 24R-hydroxylases are two analogous enzymes which utilize the vitamin as a common substrate for the catalytic production of 1 alpha,25-dihydroxyvitamin D3 and 24R,25-dihydroxyvitamin D3. These two enzymes are complexes of NADPH-ferredoxin reductase, and an (Fe-S)-cluster containing ferredoxin with a redox potential that allows the ultimate transfer of reducing equivalents to the terminal oxidases distinctly known as cytochromes P-450(1) alpha and P-450(24). We have used in vitro immunizations of splenocytes obtained from mice sensitized with the purified cytochrome P-450(1) alpha to generate three hybridoma clones from fusion with p3 x 63.Ag8.653 myeloma ATCC cells which selectively secrete monoclonal antibodies (MAbs) of the IgM class. The MAbs have been partially purified by ammonium sulfate fractionation followed by size separation chromatography on Sephacryl S-200-HR. We have compared the structural similarities and differences between the two kidney enzymes in Western Blot analyses using horseradish peroxidase conjugated goat anti-mouse Igs specific for the heavy and light chains of mouse IgA, IgG and IgM. The MAbs from all three clones recognized and interacted with apparent common epitopes of the two hydroxylases but selectively discriminated against liver microsomal P-450LM2 type and adrenal mitochondrial P-450SCC cytochromes. The cytochromes P-450(1) alpha and P-450(24) were detected as two separate bands with approximate molecular weights of 57 and 55 KDa, respectively. In reconstitution of hydroxylase activities in vitro, the MAbs were equally effective in inhibiting the 1 alpha-hydroxylation and 24R-hydroxylation reactions. The ratio of micrograms of Igs to pmol cytochrome P-450 for a 50% inhibition of either activity was approximately 25. These results, collectively, seem to suggest the existence of a precursor-product relationship between the kidney mitochondrial 1 alpha- and the 24R-hydroxylases, or perhaps, a common ancestral origin. Immunochemical peroxidase anti-peroxidase staining of kidney tissue first exposed to the MAbs revealed that only the proximal tubular segment of the nephron was specifically enriched with the cytochromes.
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PMID:Monoclonal antibodies to chick renal calcium regulating hemeproteins. Biochemical and immunohistochemical interactions with mitochondrial 25-hydroxyvitamin D3 hydroxylases. 172 40

Spleen cells from mice immunized with human chorionic gonadotropin (hCG) were fused with mouse myeloma cells and four hybridomas, secreting monoclonal antibodies, were selected for further studies. In cross-check tests against tissue extracts and peptide hormones it was established that mAb IB10 (IgG1) reacted positively against hCG only but not with other hormones. Ascites from this hybridoma was fractionated by ammonium sulphate precipitation and by FPLC to isolate the IgG fraction. Subsequently, the purified mAb was conjugated with horseradish peroxidase and used for development of a simple test for pregnancy diagnosis. Some preliminary experiments have shown that the test is very specific and reproducible.
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PMID:Production and application of monoclonal antibody specific for human chorionic gonadotropin. 219 23

A case of monoclonal IgE type lambda with IgE levels about 1 mg/ml has been followed for 6 years. Except for a slight asthma no signs of malignancies, parasitic infestations or other known diseases compatible with hyper-IgE have been found. By the combination of fractional ammonium sulphate precipitation, gel filtration, chromatofocusing, and subtraction affinity chromatography the IgE protein was isolated in an immunochemically pure and homogeneous form. Immunofluorescence of bone marrow cells showed about 1% IgE plasma cells. The amount of basophil bound IgE was 42 ng/10(6) cells, and histamine release from basophils challenged with anti-IgE was not different from that in atopic control persons, indicating a within-allergic-patients normal amount of IgE receptors. The protein-A reactivity was 0.4% equivalent to well-known IgE myeloma proteins. No antigen specificity of the IgE protein was found. Only a few cases of asymptomatic hyper-IgE are known, and it cannot be ruled out that this represents a premyeloma condition, since a similar case terminated in a malignant lymphoma.
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PMID:Clinical and immunological aspects of a case of monoclonal hyper-IgE. Isolation of IgE-protein, estimation of basophil cell bound IgE and histamine release. 240 96

Monoclonal antibodies to different domains of the porcine intestinal 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] receptor have been produced. A nuclear extract enriched in the 1,25-(OH)2D3 receptor was prepared from small intestinal mucosa of young pigs. The receptor was purified an additional 6600-fold by chromatography on DNA-cellulose, ammonium sulfate precipitation, gel filtration high-performance liquid chromatography, and DEAE-Sepharose chromatography, with an overall yield of 23% and an average purity of 24%. A BALB/c mouse immunized with this material developed serum polyclonal antibodies to the 1,25-(OH)2D3 receptor, as demonstrated by a change in sedimentation of the porcine receptor on sucrose gradients. Spleen cells from this animal were fused with mouse myeloma cells (P3-NSI/1-Ag4-1, SP2/0-Ag14), and 24 hybridomas secreting antibodies to the 1,25-(OH)2D3 receptor were identified by both a radiometric immunosorbent assay and an immunoprecipitation assay. Twenty-one hybridoma lines were cloned by limiting dilution and further characterized as subclass IgG1 antibodies with the exception of one which is an IgA. All but two of the antibodies cross-react with the 1,25-(OH)2D3 receptor from both mammalian (human, monkey, and rat) and avian (chicken) intestine; two antibodies recognize only porcine intestinal receptor. All antibodies are unreactive to the vitamin D serum transport protein. Eight of the antibodies bind denatured receptor on an immunoblot. A solid-phase competition assay was used to identify four groups of antibodies that bind to distinct epitopes on the 1,25-(OH)2D3 receptor. One antibody from each of the four groups was used to examine the effect of antibody binding on the DNA-binding activity of the receptor-hormone complex. One antibody completely inhibited the binding of the 1,25-(OH)2D3 receptor complex to DNA-cellulose, suggesting that the epitope for this antibody may be located in the polynucleotide binding domain of the protein. Antibodies from two additional groups only slightly perturbed DNA binding, while one had no effect, suggesting that these antibodies bind to receptor epitopes distant from the region of the polypeptide directly involved in polynucleotide binding. These antibodies that are directed to several different binding sites on the 1,25-(OH)2D3 receptor provide important new tools to probe the biochemistry and topology of the 1,25-(OH)2D3 receptor and to investigate its role in mediating target tissue response to hormone.
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PMID:Monoclonal antibodies to the porcine intestinal receptor for 1,25-dihydroxyvitamin D3: interaction with distinct receptor domains. 242 89

Rabbit uterine collagenase was purified from the medium of involuting uterus (1-2 days postpartum) in culture using ammonium sulfate fractionation, DEAE-cellulose, heparin-affinity, and high performance liquid chromatography. The enzyme was purified more than 1600 fold. Hybridoma cell-lines producing monoclonal antibodies were prepared by fusing the spleen cells of mice immunized with the purified enzyme with mouse myeloma cells (Sp2/O-Ag14). The hybridoma cells were selected with HAT medium, cloned, and screened by ELISA. Antibody-producing ascites were prepared by injecting hybridoma cell-lines into the peritoneal cavities of mice. Western-blot analysis indicated that the antibodies recognized a polypeptide having a molecular weight of 52,000. The IgG isolated from the ascites inhibited the enzyme. Indirect immunofluorescent staining demonstrated that polymorphonuclear leukocytes (PMNs) in the superficial layer of alkali-burned corneas contained collagenase, whereas stromal cells and PMNs within the stroma were not stained by the antibodies. Our results suggest that collagenases produced by rabbit PMNs are different from those produced by fibroblasts from cornea. We hypothesize that PMNs in alkali-burned corneas secrete all or most of their collagenases by degranulation at the anterior surface of the cornea, and then continue to migrate into the deeper portion of the stroma.
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PMID:Development of monoclonal antibodies recognizing collagenase from rabbit PMN; the presence of this enzyme in ulcerating corneas. 243 Jul 58

Splenocytes from BALB/c mouse immunized with potato virus X (PVX) and myeloma cells (SP/2) were fused. After cloning and three screening cycles, three hybridoma cell lines secreting strain-specific monoclonal antibodies (MCAs) against PVX were obtained. These cell lines were stable for 20 generations and after storage in frozen form (in liquid nitrogen) for one year. The chromosome numbers of the three hybridoma cell lines clustered in the 92-102 range. The MCAs all belonged to the IgG3 immunoglobulin subclass. The medium supernatant antibody titer detected by indirect ELISA was 1:320-1:640. The mouse ascites antibody titer was 1:102,400-1:204,800, which was more than 300 times the rabbit antiserum titer (1:320). The MCAs had a neutralizing effect on the antigen, with neutralization titer of 1:10(2). There were no apparent changes in antibody activity after repeated freezing/thawing cycles, ammonium sulfate precipitation, or freeze-drying.
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PMID:Establishment of hybridoma cell lines secreting MCAs against potato virus X and determination of their physical and chemical characteristics. 249 19

The superoxide-forming NADPH oxidase of resting macrophages can be activated in a cell-free system by certain anionic amphiphiles, most notably SDS. Activation requires the cooperation of membrane-associated and cytosolic components. We now report that at least two cytosolic factors are required for SDS-elicited activation of NADPH oxidase of guinea pig macrophages. Treatment of cytosol with ammonium sulfate at 37% saturation led to the partition of the two factors in the supernatant and precipitate fractions (termed components sigma 1 and sigma 2, respectively). Although each fraction by itself was inactive, recombining them resulted in complete recovery of the original ability of native cytosol to support SDS-elicited superoxide production by octyl-glucoside solubilized macrophage membranes. Both components are proteins, as shown by their susceptibility to trypsin and proteinase K, and were inactivated by heating at 60 degrees C. sigma 2, but not sigma 1, was inactivated by treatment with the covalent sulfhydryl reagent N-ethylmaleimide. On high-performance gel filtration, sigma 1 was found to have a molecular mass of 30 to 52 kDa, whereas sigma 2 eluted with molecules of 150 to 440 kDa. Component sigma 1 was partially purified from the ammonium sulfate supernatant fraction of cytosol by hydrophobic interaction chromatography followed by gel filtration. A material behaving like sigma 1 was also found to be present in the cytosol of guinea pig thymus cells, lymph node lymphocytes and brain and of the mouse myeloma cell line MOPC 315. However, sigma 2 appears to be strictly phagocyte specific. The molecular characteristics of sigma 1 components from nonphagocytic cells were similar to those of macrophage sigma 1, as shown by their presence in the supernatant, after treatment of cytosol with ammonium sulfate at 37% saturation, a molecular mass close to 30 to 52 kDa and a similar behavior on hydrophobic interaction chromatography. These findings raise the possibility that cytosolic component sigma 1 might be the bearer of a cellular function, more general than the one suggested by its role in the activation of NADPH oxidase of phagocytes.
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PMID:Activation of the superoxide-forming NADPH oxidase of macrophages requires two cytosolic components--one of them is also present in certain nonphagocytic cells. 255 80

There are inherent technical difficulties in measuring IgG rheumatoid factor (IgG-RF) in the serum of patients with rheumatoid arthritis (RA). These arise from measuring a reaction between two IgG molecules and the interference of IgM-RF in the reaction. We compared the prevalence of IgG-RF in whole sera and purified IgG fractions from 58 RA patients (43 of whom were latex or sheep cell agglutination positive). Methods of purification were: ammonium sulphate precipitation and DEAE cellulose or protein A-Sepharose chromatography. IgG-RF was measured by two methods: (1) radioimmunoassay and ELISA with a monoclonal myeloma IgG (IgG4,K) as the antigen and radiolabelled rabbit anti-human IgG (previously absorbed on a column with IgG4,K) as the second antibody; (2) ELISA using rabbit IgG as the antigen and a peroxidase conjugated goat anti-human IgG as the second antibody. When whole sera were assayed, 18 (31%) contained IgG-RF. In contrast, only three of the IgG fractions (5%) were positive for IgG-RF by all methods, while the remainder were uniformly negative. These results suggest that IgG-RF determination in whole sera does not accurately reflect IgG-RF activity.
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PMID:IgG rheumatoid factor in purified IgG fractions and whole sera from patients with rheumatoid arthritis. 273 Sep 84


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