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  • 1
    ISSN: 1573-8280
    Keywords: serodiagnosis ; human monoclonal antibody ; human-human hybridoma ; porcine pancreas ; porcine antigen ; lung cancer and ovary cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Antigens, recognized by human monoclonal antibody (HB4C5) generated from a lung cancer patient, were found to occur in porcine pancreas. The antigens-I and -I1 were purified from crude trypsin of porcine pancreas, only by Mono Q column chromatography, and were eluted at 260 and 300 mM NaCl in 10 mM Tris-HCI buffer, pH 7.4, respectively. These antigens differed from trypsin in molecular weight, elution pattern from the Mono Q column, and their reactivity with HB4C5. The molecular weights of the two antigens were almost the same at around 35000. These were used for serodiagnosis with an assay system based on 96-well immunoplates. The reactivities of antigens-I and -II with various sera were similar. When the reactivity of IgG in serum with antigen-II was measured, absorbance at 415 nm in the case of normal and lung cancer patients was 0.178 ± 0.056 and 0.492 ± 0.136 (p 〈 0.005). The rates of positive reaction in ovary, larynx, uterus, lung and liver cancers were more than 50%, but the rates in stomach and breast cancers were less than 30%. Positive reaction was hardly detected in pancreas cancer and normal controls.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-0778
    Keywords: lymphocytic clonal growth factor ; human-human hybridoma ; low cell density ; lymphocytic cell lines ; and adhesive cell lines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human-human hybridoma SH-76 cells were found to produce a factor that supported the growth of lymphocytic cells at low densities. The factor was purified from serum-free conditioned medium of the hybridoma cells by a successive application of ammonium sulfate precipitation, DEAE-Toyopearl, TSK G3000 SW and DEAE-5PW column chromatograph. The purified factor was a 72K single protein. The factor showed marked growth stimulating effect on lymphocytic cell lines, but had no effect on the growth of human adhesive cancer cell lines. Thus, the factor is a lymphocytic clonal growth factor (LCGF), as found previously in human plasma (Miyata, 1988). The LCGF of SH-76 cells could be produced in growth factor-free RPMI medium and purified easily from the conditioned medium. The factor is inactivated by heating at over 80°C, but is much more stable than the LCGF in human plasma.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-0778
    Keywords: lymphocytic clonal growth factor ; human-human hybridoma ; human plasma ; low cell density and lymphocytic cell lines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human plasma was found to contain a macromolecular protein which can grow even a single cell of human lymphocytic cell lines (B-lymphoblastoid cell line HO-323-3 and T-lymphoblastoid cell line CCRF-CEM) and human-human hybridoma clones (SH-9, SU-1 and HB4C5) in a dish, but it has no effect on the growth of epithelial cell lines (lung cancer cell lines PC-8, QG-56 and QG-90). The proliferating activity for lymphocytic cell lines was gradually decreased at 4 or -20°C and dramatically decreased by heating at more than 60°C for 15 min. From human plasma, active fractions were purified by a successive application of Ca2+ treatment, ammonium sulfate fractionation, DEAE-5PW column chromatography (FPLC) at pH 7.6. These active fractions were divided into at least three proteins by DEAE-5PW chromatography at pH 8.5 and chromatofocusing. These purified factors, named lymphocytic clonal growth factors (LCGFs), had similar molecular weights of about 600 K and each factor consisted of a 180 K and two 210 K subunits associated with hydrogen bondings. By the addition of 5 μg/ml of each factor into culture media, incidences of human-human hybridomas and cloning efficiencies of the hybridomas increased several-fold.
    Type of Medium: Electronic Resource
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