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)

Hybridomas producing proinsulin antibodies were cloned by limiting dilution of cell cultures obtained by fusion of splenocytes of immunized mice with immortal myeloma cells. Some proinsulin monoclonal antibodies crossreacted with labelled insulin but none did with labelled C-peptide indicating that the involved epitopes were at one of the insulin/C-peptide junctions or included in the insulin moiety. Hybridoma supernatants were assayed for IgG concentration by a solid phase assay and for ligand binding by a radiobinding assay and an enzyme linked immunosorbent assay. The half-life of immune complexes formed with radioligand was measured and, as expected, correlated with affinity as measured by the method of Scatchard. Antibody titres determined by enzyme linked immunosorbent assay did not correlate to those measured by radiobinding assay. IgG concentration correlated to enzyme linked immunosorbent assay titres but not to radiobinding assay titres. Finally, a significant correlation was found between radiobinding assay titre and the product of enzyme linked immunosorbent assay titre by the period of immune complexes. It is concluded that, except for very low affinity antibodies, enzyme linked immunosorbent assay is a capacity assay whereas radiobinding assay is influenced by both antibody concentration and affinity. The former assay is thus best suited to detecting low affinity antibodies whereas the latter is more efficient in the presence of low levels of high affinity antibodies.
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PMID:Influence of affinity of antibodies upon their detection by liquid phase radiobinding assay and solid phase enzyme linked immunosorbent assay. Demonstration using monoclonal antibodies raised against rDNA human proinsulin. 191 50

Two hybridomas producing monoclonal antibodies against mouse beta nerve growth factor (NGF) were obtained from the fusion of hyperimmune splenocytes from rats immunized with polymerized beta-NGF and Sp2/0.Ag mouse myeloma cells. The monoclonal antibodies coded IgG 24 and 30 produced and secreted by the hybrid cells are both of the IgG2a subclass. Both monoclonal antibodies are capable of recognizing native NGF coated on microassay plates as well as the denatured factor on Western blots. However, only IgG 30 has been found to block NGF-induced process outgrowth from the rat pheochromocytoma cell line (PC12) as well as NGF-induced increase in choline acetyltransferase activity in rat primary septal cell cultures. In addition, only IgG 30 was able to detect immunocytochemically NGF-immunoreactive sites in fixed tissue. And, finally, IgG 24 could not compete for IgG 30 binding to immobilized native NGF. Consequently, it appears that these antibodies are recognizing different epitopes on the NGF molecule. Neither monoclonal antibody displayed any crossreactivity with serum albumin, aprotinin, epidermal growth factor or insulin. A hybrid-hybridoma producing bi-specific anti-NGF anti-horseradish peroxidase (HRP) monoclonal antibodies was generated from the fusion of an azaguanine resistant anti-HRP hybridoma, coded RAP2.Ag and the anti-NGF IgG 30 hybridoma treated with emetine. The potential merits of using these bi-specific antibodies in combination with their mono-specific anti-NGF parent in a homogeneous sandwich immunoassay for the quantitation of NGF are discussed.
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PMID:Two distinct monoclonal antibodies raised against mouse beta nerve growth factor. Generation of bi-specific anti-nerve growth factor anti-horseradish peroxidase antibodies for use in a homogeneous enzyme immunoassay. 199 53

Novel islet cell, duct cell, and acinar cell markers have been identified by monoclonal autoantibodies (Maab) derived from prediabetic BB rats. Spleen cells from two rats that both developed diabetes after splenectomy were fused with mouse myeloma cells. A cellular immunoradiometric assay for differential reactivity toward the surface of two closely related, insulin- and non-insulin-producing rat islet tumor cell lines was used to select and clone several IgM-producing hybridomas. The supernatants were finally characterized by two-color immunofluorescence with islet hormone antisera on frozen sections of human, monkey, and rat pancreas. Maab EB52 stained PP cells, but also few A cells on rat pancreas. Maab CA812 identified a subpopulation of islet D cells on rat, human and monkey pancreas. Although the CA812-reactive antigen and somatostatin were coexpressed in most D cells in adult rat pancreas, only a few islet D cells were stained in the newborn pancreas. The CA812-reactive antigen was not detected in somatostatin-producing cells in the duct epithelium. Maab H37 and IF5 selectively stained acinar cells in rat, human, and monkey pancreas, whereas Maab DA39 identified the rat ductal epithelium including the scattered endocrine cells of the ducts. In summary, B lymphocytes producing autoantibodies to pancreatic endocrine, exocrine, and ductal markers are present in prediabetic BB rats and can be detected by use of transformed pluripotent islet cells as target. Such B lymphocytes can be immortalized to produce monoclonal antibodies to study their role in insulin-dependent diabetes mellitus pathogenesis and to clarify the development of the pancreas.
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PMID:Novel islet, duct, and acinar cell markers defined by monoclonal autoantibodies from prediabetic BB rats. 212 46

Multiple myeloma associated with sclerotic bone lesions and polyneuropathy represents a distinct subset of the plasma cell dyscrasias. We describe a case of biclonal gammopathy (the second case reported), insulin-resistant diabetes mellitus, and no evidence for anti-insulin receptor antibodies. After treatment with chemotherapy and irradiation, the diabetes resolved, the polyneuropathy lessened greatly, and the patient is alive without evidence of progression five years later. The reports of 95 other cases are reviewed. This syndrome occurs in younger patients (mean age, 48 years) and is frequently associated with organomegaly, endocrinopathies, and skin changes. Irradiation to the sclerotic bone lesions frequently lessens the neuropathy and endocrinopathies and may result in long-term remission. The mechanism of action leading to the systemic effects seen in this syndrome is unknown but is likely related to proteins secreted by the abnormal plasma cells.
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PMID:Syndrome of plasma cell dyscrasia, polyneuropathy, and diabetes mellitus. 218 97

The BB rat provides an excellent animal model for type 1 (insulin-dependent) diabetes mellitus. Cytotoxic autoantibodies against pancreatic beta cells have been found in the sera of both patients with type 1 (insulin-dependent) diabetes and BB rats. These antibodies have been implicated in the pathogenesis of the disease. In this study, a monoclonal autoantibody, designated KT1, has been developed by the fusion of spleen cells from a BB rat and a mouse myeloma cell line. KT1 was found to be of the immunoglobulin M isotype and reacted specifically with islet cells. In microcytotoxicity assays KT1 was shown to mediate complement-dependent lysis of approximately 30% of a rat insulinoma cell line and 25% of rat pancreatic islet cells in culture. It did not cause lysis of the other cell lines tested. KT1 has been demonstrated, by indirect immunofluorescence, to bind specifically to a cell surface antigen on live and acetone-fixed islet cell cultures from Wistar rat neonates and to rat insulinoma cells. Western blotting experiments revealed reaction to a 68-kDa protein from rat insulinoma cell extracts. This monoclonal antibody may have clinical relevance as it exhibits properties similar to the islet cell surface antibodies present in the sera of BB rats.
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PMID:A cytotoxic monoclonal autoantibody from the BB rat which binds an islet cell surface protein. 240 25

An unlimited supply of suitable antisera is wanted for immunoassays, analysis of antigenic determinants and precise localization of antigens in biological systems. Therefore, we produced a monoclonal antiporcine insulin antibody by the hybridoma technology and assessed it in comparison with polyclonal antibody. The spleen cells of BALB/c mice immunized against porcine insulin were hybridized with mouse myeloma cells (P3-X63-Ag8-U1). The monoclonal antibody thus generated was shown to have high binding capacity and specificity to porcine insulin in radioimmunoassay. It reacted with human insulin as well, but did not crossreact with other polypeptide hormones produced in the pancreatic islets such as glucagon, somatostatin and pancreatic polypeptide. In immunohistochemistry human and dog islets were stained by this monoclonal antibody. Rat islets were not stained, although they reacted with polyclonal anti-insulin antibody. The insulin of human serum samples measured using the monoclonal antibody was tightly correlated with that using the polyclonal antibody. These observations indicate that our hybridoma-derived monoclonal antibody is useful for immunoassay as well as localization of insulin.
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PMID:Production of anti-insulin monoclonal antibody and its application to immunoassay of insulin and immunohistochemistry. 246 89

By comparison, it was shown that ascaris protein as carrier was far more effective than BSA and mouse serum albumin to make human c-peptide (HCP) immunogenicity in mice. Twelve monoclonal antibodies (McAb) were obtained after fusion of NS-1 myeloma cells with spleen cells from two BALB/c mice immunized with HCP-ascaris protein conjugate. Eight McAb were characterized and all of them were of IgG1 subclass. Affinities of the McAb for HCP were in the range of 0.03-0.85 X 10(9) mol-1. Although all 8 McAb reacted with both HCP and human proinsulin (HPI), they could specifically recognize human insulin from HPI and were highly species specific without cross-reaction with proinsulin, insulin and c-peptide of other species tested. Through analysis of binding with different HCP fragments, their antigenic determinants were located at either N or C terminal of HCP. The results of plasma HCP radioimmunoassay in 20 normal fasting human adults showed that the value (0.47 +/- 0.16 pmol/ml) determined by rabbit PcAb (R2303) against BSA-HCP was significantly higher than that (0.29 +/- 0.13 pmol/ml) determined by McAb-1.
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PMID:[Studies on monoclonal antibodies directed against human C-peptide]. 247 69

Monoclonal antibodies were raised against pregnancy-associated endometrial alpha 1-globulin (alpha 1-PEG), a 32 KD insulin-like growth factor binding protein (IGF-BP), which represents a major secretory product of the human decidualized endometrium during pregnancy. This class of IGF-BP has been implicated in the modulation of action, inhibitory and stimulatory, of insulin-like growth factors. Immunization with the protein purified from pregnancy endometrium resulted after myeloma fusion in the isolation of six hybridoma clones and the antibodies produced were characterized. The Ka of the antibodies ranged between 4.75 x 10(9) M-1 and 0.7 x 10(8) M-1. In Western blots all monoclonal antibodies reacted with purified protein of molecular weight 32 KD and specifically detected this IGF-BP species in culture medium and cytosolic extracts of pregnancy endometrium and amniotic fluid. The monoclonal antibodies appear to define three epitope-bearing regions as evidenced by their reactivity to polypeptide fragments of the protein. After synthesis and secretion by tissue explants in vitro the protein is susceptible to cleavage into fragments possessing different monoclonal antibody-defined reactivity. Employing immunohistochemical techniques the protein was principally localized to decidual cells in tissue sections of pregnancy endometrium and solely to these cells after enzymic digestion of the tissue. The implications of these results are discussed with respect to potential role of IGF-BP in the action of IGF upon the IGF-1 receptor-bearing populations, including lymphocytes and trophoblast cells, D in the decidua.
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PMID:Monoclonal antibodies to human secretory "pregnancy-associated endometrial alpha 1-globulin," an insulin-like growth factor binding protein: characterization and use in radioimmunoassay, Western blots, and immunohistochemistry. 248 Jan 40

A radioimmunoassay for glycated serum protein (GSP) was developed using monoclonal antibody to glucitollysine and polystyrene beads coated with Coomassie-Brilliant-Blue (CBB) as adsorbent for serum protein. The monoclonal antibody was raised by immunizing BALB/c mice with reduced glycated LDL and fusing their spleen cells with mouse myeloma cells. CBB-coated polystyrene beads were introduced to absorb a constant amount of serum protein. The protein adsorbed on the CBB-coated beads was reduced by NaHB4, and after treatment with radiolabeled antibody, the radioactivity of each bead was counted with an automatic gamma-counter. The standard glycated protein used was reduced glycated human serum albumin, in which 8 of 59 lysine residues were glycated. The intra- and interassay coefficients of variation of GSP were 4.8-6.5% and 1.6-6.0%, respectively. The GSP level of diabetic patients was significantly higher than that of normal controls (1.97 +/- 1.23 vs. 0.47 +/- 0.21 nmol/mg-protein; mean +/- SD, p less than 0.001). The GSP levels of patients with insulin-dependent and non-insulin-dependent diabetes mellitus were 3.03 +/- 1.05 and 1.51 +/- 1.00 nmol/mg-protein, respectively. A good correlation was found between the levels of GSP and hemoglobin A1c (HbA1c) (r = 0.85, p less than 0.001). In patients admitted to the hospital for diabetes education and glycemic control, the GSP level decreased 43 +/- 12% with the decrease in the fasting plasma glucose level (39 +/- 13%) and the mean daily plasma glucose level (MPG, 47 +/- 15%) in a four week period after admission, whereas the HbA1c level decreased only 13 +/- 6% during this period.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Radioimmunoassay of glycated serum protein using monoclonal antibody to glucitollysine and coomassie-brilliant-blue-coated polystyrene beads. 251 33

Monoclonal antibodies (MAbs), for human use require chemical and biological purity. The best approach seems in vitro cultivation in a serum-, protein-free medium. A basal defined culture medium has been developed to sustain optimal hybridoma cell growth and MAb secretion. It consists of Iscove's Dulbecco's modified, Eagle's, Ham's F12 and NCTC 135 media in a 5:5:1 mixture (v/v/v), to which glucose is added to reach a final concentration of 25 mM, glutamine to 4-6 mM, 2-mercaptoethanol to 50 microM, Pluronic F68 to 0.01-0.1% (w/v), Hepes to 25 mM and NaHCO3 to 3 g/l. Hybridoma cells, derived from Sp 2/0 myeloma and secreting a MAb to a human milk fat globule membrane-associated high molecular weight glycoprotein, were cloned in this medium containing 1% (v/v) fetal calf serum and then sequentially adapted in serum-free medium further supplemented with transferrin and insulin, both at 10 micrograms/ml. Clones producing immunoreactive MAbs secrete a mean of 50 micrograms IgG/ml, i.e., ca. 80% of the concentration reached in Dulbecco's modified Eagle's medium containing 10% serum. When cells were cultured in spinner flasks with a semi-continuous mode of cultivation (with a daily removal of 20% of the volume and its replacement by fresh culture medium), in serum-free medium further supplemented with 10 nM estradiol, a mixture of trace elements and albumin (at 30 micrograms/ml) complexed to linoleic acid, MAb secretion reached 100 micrograms/ml and became equal or higher to that obtained in serum-containing medium. MAb secretion was not decreased and was even significantly increased during the growth phase, when transferrin was replaced by another iron source, i.e., ferric citrate at 500 microM associated with 20 microM ascorbic acid. Finally, deletion of insulin and of albumin-linoleic acid did not affect significantly cell density nor MAb secretion. In conclusion, it appears from this study that semi-continuous cultivation in spinner flasks of hybridoma cells, after cloning and progressive adaptation, in a chemically defined, serum- and protein-free medium, permitted MAb secretion to be increased to a mean of 144 micrograms/ml, i.e., multiplied by a factor of ca. 1.5 compared to culture of these cells in serum-containing medium under the same conditions and by a factor of ca. 2.4 compared to cultivation in serum-containing medium in flasks.
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PMID:Optimisation of hybridoma cell growth and monoclonal antibody secretion in a chemically defined, serum- and protein-free culture medium. 264 56


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