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)

Like other small-sized neurotransmitter molecules, glutamate (Glu) was conjugated to carrier proteins via glutaraldehyde (G). Human serum albumin (HSA) and thyroglobulin (TH) conjugates were alternately injected into mice. When a relevant immune response was obtained for antibody affinity and specificity, hybridization of spleen activated lymphocytes with SP2/O/Ag myeloma cells was performed. Supernatant culture media of hybridomas were tested for the presence of anti-conjugated Glu antibodies with our ELISA method. Selected hybridomas giving good antibody affinity and specificity were then cloned by the limiting dilution technique. Using DEAE-chromatographed ascites fluid, Glu reactivity was observed on the cortex and the hippocampus. Staining obtained with this monoclonal antibody was in agreement with that observed with previous polyclonal antisera directed against conjugated Glu or monoclonal anti-gamma-glutamyl-Glu antibody.
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PMID:Monoclonal antibody directed against glutaraldehyde conjugated glutamate and immunocytochemical applications in the rat brain. 256 32

A monoclonal antibody directed against the N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP), named KY-ANP-II, was produced by fusion of a non-producing mouse myeloma cell line, X63-Ag8.653, with spleen cells from a BALB/c mouse immunized with synthetic alpha-human ANP (alpha-hANP) conjugated to bovine thyroglobulin. The obtained antibody showed a high affinity for alpha-hANP with a Ka of 6.6 X 10(10) M-1. With this monoclonal antibody, a specific radioimmunoassay for alpha-hANP was established. The minimal detectable level of alpha-hANP in this radioimmunoassay was 0.8 fmol (2.5 pg)/tube, and IC50 was 8 fmol (25 pg)/tube. The radioimmunoassay recognized alpha-rat ANP on an equimolar basis, whereas there was no detectable cross-reactivity with alpha-ANP [4-28] or alpha-ANP [5-28]. Thus, the monoclonal antibody can detect a hormonal form of ANP, alpha-ANP, but not neuropeptide forms of ANP, alpha-ANP [4-28] and alpha-ANP [5-28]. These results indicate that KY-ANP-II becomes a useful tool for preferential detection of a circulating form of ANP and for investigation of the physiological and pathophysiological significance of ANP as a hormone.
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PMID:A monoclonal antibody against N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP): a useful tool for preferential detection of naturally circulating ANP. 296 82

We produced three monoclonal antibodies against different epitopes of the precursor form of atrial natriuretic polypeptide (ANP), gamma-human ANP (gamma-hANP) [human atrial natriuretic factor (ANF)-(1-126)]. Monoclonal antibodies were prepared by fusion of mouse myeloma cells X63-Ag8.653 with spleen cells from BALB/c mice immunized with synthetic alpha-hANP or gamma-hANP-(1-25) [human ANF-(1-25)] conjugated to thyroglobulin. Of three monoclonal antibodies obtained, two were directed against the alpha-ANP sequence: one antibody (KY-ANP-I) recognizes the N-terminal half of the ring structure, while the other (KY-ANP-II) recognizes the N-terminal sequence of alpha-hANP. The third antibody (KY-ANP-III) was against gamma-hANP-(1-25). With the aid of rabbit polyclonal anti-alpha-ANP-(17-28), we developed sandwich enzyme immunoassays. An enzyme immunoassay for alpha-hANP using KY-ANP-I and polyclonal Fab' was very sensitive (0.01 fmol/tube) and we could detect 0.6 pg/ml of plasma alpha-hANP without extraction. Thus, monoclonal antibodies against gamma-ANP are useful tools for investigating the significance of ANP and related peptides derived from gamma-ANP.
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PMID:Preparation of monoclonal antibodies against atrial natriuretic polypeptide precursor and application to highly sensitive sandwich enzyme immunoassay. 297 70

Human thyroglobulin was used as an antigen for the development of monoclonal antibodies by the hybridoma technique. Spleen cells of BALB/c mice immunized with human thyroglobulin were fused with SP2/0 mouse myeloma cells. Seven clones secreting specific monoclonal antibodies to thyroglobulin were established. Two of these monoclonal antibodies have been purified and characterized. Their equilibrium association constants (Ka) as determined by Scatchard analysis were 0.24 X 10(11) L/M and 1.4 X 10(11) L/M respectively. The specificity of both these antibodies was validated by immunohistochemical staining of human tissues (normal human thyroid, brain, salivary gland, skeletal and smooth muscle, mucous membrane, parathyroid, adrenal) obtained from autopsy material. Only follicles and follicular cells of thyroid tissue were stained by both the monoclonal antibodies. H10 I monoclonal antibody was used for constructing a standard curve for in vitro immunoassay using a solid phase ELISA technique. The minimum amount detectable was 7.8 ng/ml. Thirty six sera from patients of various thyroid disorders were evaluated using ELISA and compared with conventional RIA. A good agreement was seen (r = 0.92) between the two techniques. These specific monoclonal antibodies may prove to be valuable for in vitro immunoassays and in vivo immunoscintigraphy.
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PMID:Monoclonal antibodies to human thyroglobulin: production and characterization. 313 Dec 34

Selectivity for the carboxy-terminus of angiotensin II (Ang II) and the high affinity of antibodies are prerequisites for clinical assays that evaluate Ang II in the presence of Ang I. A high-affinity monoclonal antibody (Kd = 7.1 X 10(-11) mol/l) was produced and used for the measurement of plasma Ang II. C3H mice were immunized with Ang II coupled by its carboxy-terminus to thyroglobulin. Somatic cell fusion between spleen cells and SP 2/0 myeloma cells, repeated subcloning and re-injection into mice yielded ascites containing sufficient antibody at a 2 X 10(7)-fold dilution. Radioassay standard curves show 50% tracer displacement when 32 fmol unlabelled Ang II is added and 2 fmol Ang II can be detected. Cross-reactivities, taking the reactivity with Ang II as 1.00 are: Ang I 0.003, Ang (1-7) 0.00001, Ang III 1.05, Ang(3-8) 0.88 and Ang(4-8) 0.75. Fast extraction of angiotensin from 2 ml plasma by reversible adsorption to phenylsilylsilica (Bondelut PH) provides recoveries of 96-102%. During angiotensin converting enzyme inhibition with 25 mg intravenous captopril, plasma immunoreactive Ang II decreased in supine normal volunteers from 8.6 +/- 3.6 to 4.5 +/- 3.4 fmol/ml (P less than 0.01, n = 8). It thus appears that plasma immunoreactive Ang II can now be measured after a simple extraction procedure by using monoclonal antibodies.
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PMID:Angiotensin II measurement with high-affinity monoclonal antibodies. 324 Dec 30

Four strains of mice were immunized with 6 different conjugates of 3-O (carboxymethyl-oximino)-18-hydroxycortisol to bovine serum albumin (3 preparations), turkey serum albumin, porcine thyroglobulin, and keyhole limpet hemocyanin. Spleens from 7 of 48 mice immunized were fused with Fox/NY and/or HL-1 Friendly myeloma cell lines, yielding many positive clones for antibody formation. Short cross-reactivities were done in 293 culture supernatants and were found to have low cross-reactivity (less than 0.001%) to cortisol, but very high cross-reactivity to 18-hydroxy-11-deoxycortisol (70 to 140%). One clone showed over 100% cross-reactivity with all the 18-hydroxylated steroids studied. The major problem encountered in the generation of monoclonal antibodies was the low antibody response in the vast majority of mice injected. Half the mice developed no measurable titer, and the clones evaluated from those that did produce antibodies cross-reacted with other 18-hydroxylated steroids. Nevertheless, the antibody developed could serve in radioimmunoassay for 18-hydroxy-11-deoxycortisol separated chromatographically from other cross-reacting steroids. This is important as no synthetic 18-hydroxy-11-deoxycortisol is available.
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PMID:The production of a monoclonal antibody to 18-hydroxycortisol and other 18-hydroxylated steroids. 345 46

A serum-free in vitro immunization method for the generation of hybridomas producing specific antibodies to an antigen is described. The method was tested with human thyroglobulin as antigen. The serum-free medium used (Yssel et al., 1984) consisted of Iscove's modification of Dulbecco's modified Eagle's medium, supplemented with albumin, transferrin, insulin, ethanolamine and linoleic, oleic and palmitic acids. An optimal response was obtained when splenocytes from BALB/c mice were cultured for 3 days in the presence of 1.5 nM thyroglobulin and thymocyte-conditioned medium prior to fusion with SP2/0 myeloma cells and seeding of the fused cells in microtitre plates. The frequency of positive wells, defined as the number of wells secreting anti-(thyroglobulin) antibodies/number of viable cells used for the fusion, was 1.6 X 10(-6) +/- 0.25 X 10(-6) (mean +/- SD; n = 4). Eight stable clones producing anti-(thyroglobulin) antibodies were isolated. One clone (3D12) produced antibodies reacting only with human thyroglobulin. The antibodies produced by the other clones reacted with human, murine and porcine thyroglobulins. Seven of the clones produced antibodies of the IgM class and one clone produced IgG. The specificity of 3D12 (IgM) for human thyroglobulin and the absence of any reactivity with murine thyroglobulin provides evidence for a primary response of splenocytes in culture to the presence of an antigen.
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PMID:Production of murine monoclonal antibodies against human thyroglobulin using an in vitro immunization procedure in serum-free medium. 348 50

In order to prepare an antibody directed at the common amino acid sequence of mammalian, avian, and fish luteinizing hormone-releasing hormones (LHRHs), C-terminal free LHRH was conjugated with bovine thyroglobulin, and was used as the antigen. A monoclonal antibody (LRH13) was obtained as an ascitic fluid by fusing the spleen cells of a BALB/c donor mouse immunized with the antigen to X63.Ag8.653 mouse myeloma cells followed by limiting dilution cloning and transplanting a positive clone to BALB/c mice. This monoclonal antibody seems to belong to IgG2b as it was eluted from protein A-Sepharose CL-4B with citrate buffer pH 3.5. competitive binding experiment using fragment peptides of LHRH indicated the binding site of LRH13 was a region around serine and tyrosine, and modification of mammalian LHRH by radioiodination caused a marked decrease in the binding activity. LRH13 has an affinity constant of 0.134 X 10(9) M-1 to native mammalian LHRH, and binds C-terminal free LHRH with a similar affinity (1.6X), however, it binds with higher affinities to N- and C-terminal free LHRH (12.9X), N-terminal free LHRH (10.4X), salmon LHRH (8.3X) and chicken LHRH-I (6.0X). Chicken LHRH-II, where tyrosine is replaced for histidine, has a lower affinity (0.3X) than that of mammalian LHRH. From its high affinity to N-, C-terminal free LHRH, LRH13 is also expected to bind possible precursor peptides of LHRH. Immunohistochemical staining of the brain sections obtained from rats, mice, chickens, Japanese quail, and rainbow trout successfully visualized cell bodies and fibers distributed from the olfactory bulb to the median eminence, indicating high LHRH specificity and wide crossreactivity in animal classes of this monoclonal antibody. With this antibody, LHRH-like immunoreactive substance in the pineal gland was also stained with fixation at neutral pH.
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PMID:Preparation of a monoclonal antibody to common amino acid sequence of LHRH and its application. 353 Jul 29

Human-human B-cell hybridomas were established using peripheral blood lymphocytes from patients with autoimmune thyroiditis and Graves' disease. Peripheral mononuclear cells (PMC), with or without mitogen prestimulation, were fused with HGPRT-negative human myeloma cell lines (Gm4672 and GM0462) using 44% polyethylene glycol. Developing hybridomas were screened by enzyme-linked immunosorbent assays (ELISAs) for human IgG and IgM and antibodies to human thyroglobulin (hTg) and microsomal antigen (M-Ag). A 125I-TSH binding inhibition assay was utilized for detecting antibodies to TSH receptor (TSH-R) protein. Hybridoma formation was observed only after prior mitogen stimulation of PMC. The amount of antibody secreted by the human-human hybridomas was highly variable (10 ng-100 micrograms/ml IgG/IgM). Nine and six-tenths percent of the hybrids secreted anti-hTg and 8.4% secreted anti-M-Ag. A 5% cloning efficiency was achieved, with detection of specific thyroid autoantibody secretion in one-third of the clones derived from positive hybridomas. Immunoglobulin secretion decreased with time and long-term stable clones were not achieved. Thyroid monoclonal autoantibodies to hTg, M-Ag, and TSH-R (IgG and IgM) detected during these studies were of a low affinity. In addition, antibodies were identified which exhibited marked specificity crossover between hTg, M-Ag, and nonthyroid antigens, suggesting the presence of recurrent epitopes. Such observations may help explain the multiplicity of thyroid autoantibodies in human thyroid disease and indicate a common defect in immunoregulation. We suggest that cross-reacting epitopes may be important in the derivation of thyroid-specific B-cell clones.
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PMID:A study of human-human hybridomas from patients with autoimmune thyroid disease. 355 36

A synthetic peptide corresponding to amino acids 1-19 of thyroglobulin was used to test the possibility of generating protein-reactive monoclonal antibodies by immunization in vitro with a synthetic peptide as antigen. Splenocytes from non-immunized Balb/c mice were cultured in serum-free medium for 3 days in the presence of thymocyte-conditioned medium and the synthetic peptide prior to fusion with SP2/0 murine myeloma cells. The synthetic peptide was used in its free form, i.e. not coupled to a protein carrier. Hybridomas secreting monoclonal antibodies reactive with the synthetic peptide were obtained after immunization in vitro with as little as 10 ng/ml of the synthetic peptide. Between 50 and 70% of the primary clones obtained in different experiments produced monoclonal antibodies also reactive with the intact protein. Six stable hybridomas were isolated; all produced antibodies of the IgM class. We conclude that immunization in vitro with a free synthetic peptide is an efficient method for the generation of monoclonal antibodies reactive with the intact protein.
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PMID:Production of monoclonal antibodies to thyroglobulin by in vitro immunization with a free synthetic peptide. 368 4


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