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 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

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

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

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

In the present paper, the results of detecting circulating antigen and/or antigen-antibody complexes by McAb against surface membrane antigen of adult Schistosoma japonicum were reported. The McAb, coded as 8SE4, was prepared by fusion of SP2/0 myeloma cells with spleen cells of the BALB/c mice immunized with the saline extract of adult S. japonicum. The 8SE4- directed antigen was proved to be located on the surface of the adult worm. After being purified by a DE52 column, 8SE4 was labelled with HRP and the conjugate (HRP-8SE4) was used in the test. For testing, the serum sample was first incubated with HRP-8SE4, then PEG (mw. 6,000) was added to precipitate the antigen-antibody complex. Upon centrifugation, OPD was added to the precipitate. Results were read by ELISA reader at 492nm. The OD value was found to be proportional to the amount of circulating antigen and/or antigen-antibody complexes. Results from 5 heavily infected (1,500-2,000 cercariae) rabbits showed that the OD values were raised significantly at the 6th week post infection, being 1.9-4.5 times higher than those before infection. The OD values of the 5 rabbits each lightly infected with 10-500 cercariae were also markedly raised 6 weeks post infection and reached the peak at the 8th week, then maintained in high levels until 11th week post infection. The worm burden of the 5 lightly infected rabbits were 4-326. No obvious correlations between OD values and worm loads were observed. The results suggested the existence of surface membrane-related antigen and/or antigen-antibody complexes in the circulation of infected rabbits.
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PMID:[Detection of circulating antigen and/or antigen-antibody complex by using McAb against surface membrane antigen of adult Schistosoma japonicum]. 209 94

Murine monoclonal antibody FEN-1 was derived by immunizing Balb/c mice with an affinity-purified endometrioid ovarian cancer-associated antigen recovered from ascites-derived immune complexes. Splenic lymphocytes from the immunized mouse were fused with the myeloma cells SP2/0-AG14 in the presence of PEG 1500. The hybrid cultures were screened for production of immunoglobulins reactive with an extract preparation of an endometrioid ovarian tumor by enzyme-linked immunosorbent assay and flow cytometry. One of the hybrids secretes a monoclonal antibody of the IgG3 subtype designated FEN-1, which reacts with 100% of endometrioid ovarian cancer containing adenoacanthoma by indirect immunoperoxidase on paraffin-embedded tissue. No detectable levels of antigen were found in squamous metaplasia associated with nonendometrioid tumors, and no reactivity occurred against endometrial adenocarcinomas, endometriosis, or normal ovary and endometrium. The antibody does not cross-react with mucinous tumors, nonepithelial tumors of the ovary, or gastrointestinal tissue. This antibody may be used as an aid in the diagnosis of nonmucinous ovarian carcinomas by immunohistology.
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PMID:Immunohistochemical characterization of a monoclonal antibody detecting an endometrioid ovarian cancer-associated antigen. 219 40

Spontaneous fusion between lymphoid and carcinoma cells in vivo has been described previously. Splenocytes from mice treated with LPS or mitogen have been reported to fuse better with myeloma cells using PEG as fusion agent than splenocytes from untreated mice. We report a phenomenon where immunization of mice with formalin treated, whole Haemophilus paragallinarum bacteria induced spontaneous fusion of splenocytes with myeloma cells in vitro, without the aid of any fusion agent. Co-immunization of mice with H. paragallinarum and an unrelated antigen (hen's egg white lysozyme), followed by co-culturing of the immune splenocytes with SP2/0 myeloma cells, yielded stable hybridoma cell lines producing anti-lysozyme antibodies. H. paragallinarum may be used in adjuvants to simplify the production of monoclonal antibodies, and the discovery of a promotional activity of a gram negative bacterium on cell fusion and hybridoma formation may shed new light on spontaneous fusion as a natural immune phenomenon in cancer.
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PMID:Spontaneous fusion between splenocytes and myeloma cells induced by bacterial immunization. 225 87

Methods for pre-selecting B lymphocytes were studied and investigated. First, biotinylated antigen was used for selecting B lymphocytes. These pre-selected B lymphocytes were then combined with biotinylated myeloma cells by adding streptavidin. The final formula of the selected B cell-myeloma cell was as follows: B cell-(antigen-biotin-strept-avidin-biotin)-myeloma cell. Then, this B cell-myeloma cell conjugate was fused by the pulsed electric field (PEF) method, which fused only those conjugated cells. The fusion efficiency obtained by this method was 3-15-times higher than that obtained by the non-specific poly(ethylene glycol) (PEG) fusion method. Second, avidin-antigen conjugate was used to select B lymphocytes. For this purpose, bifunctional cross-linkers such as N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) and m-maleimidobenzoyl N-hydroxysuccinimide (MBS) were chosen. Each reagent contains two heterofunctional groups which can make covalent bond with both Lys and Cys residues. Typical avidin-antigen conjugate is expressed as avidin-SPDP (or MBS)-antigen. Thus, final B cell-myeloma cell conjugate was B cell-antigen-SPDP (or MBS)-avidin-biotin-myeloma cell. The yield of this procedure was of the order of 10(-2). Here, we suggest that the pre-selection of B lymphocytes by biotinylated antigen or avidin-antigen conjugate is a new method of obtaining selected hybridoma cells which produce specific monoclonal antibodies against the antigen used for selecting B lymphocytes.
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PMID:Selective production of hybridoma cells: antigenic-based pre-selection of B lymphocytes for electrofusion with myeloma cells. 226 7

BALB/c mice were immunized with the purified p30 antigen. SP2/0 myeloma cells and immune spleen cells were fused with 50% PEG (Sigma, MW 3,350-4,000). The cell fusion rate was 93.95%, and the antibody producing rate 21.01%. The technique of limiting dilution was used for cloning of the hybridoma cells. Two hybridoma cell lines E8 and G1 secreting McAb against p30 antigen were obtained. The number of chromosome of both E8 and G1 cell lines was 98.7 +/- 6.54 and 97.7 +/- 7.77, respectively. Results of the PAGE of the ascites generated by the hybridoma cell lines E8 and G1 showed a thick protein band at the gamma-region which was absent from the ascites generated by the SP 2/0 myeloma cells. The immunoglobulin of the McAb E8 and G1 belonged to IgG2 subclass. The results of the immunohistochemical method using the horseradish peroxidase conjugated antibody showed that both E8 and G1 McAb only reacted with epithelial cells of the normal human prostatic glands and their ducts, but did not cross-react with other thirty-four different kinds of normal human tissues. The results of inhibiting ELISA test showed that both E8 and G1 McAb were only inhibited by the human seminal plasma and p30, weakly inhibited by the adult male's urine, but were not inhibited by other eight different kinds of human body fluids and secretions as well as semen from eight different species of animals. It was concluded that McAb of E8 and G1 were organ and species specific.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Establishment of hybridoma cell lines producing monoclonal antibodies against-p30 antigen]. 236 36


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