• The Journal of the Korean Society for Microbiology
• /
• v.22 no.3
• /
• pp.197-208
• /
• 1987

Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

• Annals of Clinical Neurophysiology
• /
• v.25 no.2
• /
• pp.84-92
• /
• 2023

Background: Clinical spectrum of immunoglobulin M (IgM) monoclonal gammopathy varies from IgM monoclonal gammopathy of unknown significance (IgM-MGUS) to hematological malignancies. We evaluated the clinical features, electrophysiological characteristics, and prognosis of patients with peripheral neuropathy associated with IgM monoclonal gammopathy (PN-IgM MG). Methods: We retrospectively evaluated 25 patients with PN-IgM MG. Peripheral neuropathy was classified as axonal, demyelinating, or undetermined, based on electrophysiological studies. We classified the enrolled patients into the IgM-MGUS and malignancy groups, and compared the clinical and electrophysiological features between the groups. Results: Fifteen patients had IgM-MGUS and 10 had hematologic malignancies (Waldenström's macroglobulinemia: two and B-cell non-Hodgkin's lymphoma: eight). In the electrophysiological evaluation, the nerve conduction study (NCS) criteria for demyelination were met in 86.7% of the IgM-MGUS group and 10.0% of the malignancy group. In particular, the distal latencies of the motor NCS in the IgM-MGUS group were significantly prolonged compared to those in the malignancy group (median, 9.1 ± 5.1 [IgM-MGUS], 4.2 ± 1.3 [malignancy], p = 0.003; ulnar, 5.4 ± 1.9 [IgM-MGUS], 2.9 ± 0.9 [malignancy], p = 0.001; fibular, 9.3 ± 5.1 [IgM-MGUS], 3.8 ± 0.3 [malignancy], p = 0.01; P-posterior tibial, 8.3 ± 5.4 [IgM-MGUS], 4.4 ± 1.0 [malignancy], p = 0.04). Overall treatment responses were significantly worse in the malignancy group than in the IgM-MGUS group (p = 0.004), and the modified Rankin Scale score at the last visit was higher in the malignancy group than in the IgM-MGUS group (2.0 ± 1.1 [IgM-MGUS], 4.2 ± 1.7 [malignancy], p = 0.001), although there was no significant difference at the initial assessment. Conclusions: The risk of hematological malignancy should be carefully assessed in patients with PN-IgM MG without electrophysiological demyelination features.

• Journal of Microbiology
• /
• v.45 no.6
• /
• pp.547-552
• /
• 2007

Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.8 no.2
• /
• pp.59-63
• /
• 2003

The protein disulfide isomerase (PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody (Mab) refolding and assembly which accompanies disulfide bend formation. The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb in-termediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant fur a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.1 no.1
• /
• pp.13-17
• /
• 1996

The protein disulfide isomerase(PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody(MAb) refolding and assembly which accompanies disulfide bond formation The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb intermediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant for a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.

• KSBB Journal
• /
• v.11 no.3
• /
• pp.329-339
• /
• 1996

The effects of ammonium ion on cell growth kinetics, monoclonal antibody productivity, and cell metabolism of hybridoma cells were investigated. The mouse-mouse hybridoma cell line VlIIH-8 producing mouse IgG2a was used as a model system. Ammonium ion showed an inhibitory effect on cell growth and monoclonal antibody production. New immobilized adsorbents were developed for the reduction of the inhibitory effect of ammonium ion. The ammonium ion selective zeolite, Phillipsite-Gismondine was entrapped in calcium alginate bead or in dialysis membrane and applied to the hybridoma cell culture system for the in situ removal of ammonium ion from culture media. The effects of ammonium the both serum supplemented and serum free media on the cell growth were studied by applying immobilized adsorbents of calcium alginate bead type. The results demonstrated a substantial enhancement in cell growth. Applying immobilized adsorbents of dialysis membrane type to serum supplemented media also resulted in the stimulation of cell growth, cell viability and monoclonal antibody production.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.4
• /
• pp.449-454
• /
• 2000

The heavy chain monoclonal antibody (HC MAb) was produced in suspension cultures of genetically modified Nicotiana tabacum. The HC MAb secreted to the medium was unstable due to unfavorable interactions in the plant cell medium. The addition of gelatin (5g/l) stabilized the extracellular HC MAb and increased its production 10-fold. A kinetic model was developed describing the interaction between the secretedprotein and the stabilizer. The model accounted for the inactivation of the protein by simple aggregation and general instability. It was assumed that the secreted protein and the stabilizer form a stable complex. Culturing the cells semicontinuously could further increase the productivity of HC MAb.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.5
• /
• pp.732-737
• /
• 2003

Anti-antibodies against three mouse monoclonal antibodies viz. IIB5D6, VIA6E8 and VIC1F9 (specific to bovine growth hormone) in rabbits have been generated and characterized. Ammonium sulfate fractionated and affinity-purified monoclonal antibodies were used for producing anti-antibodies. The generated anti-antibodies were against common as well as uncommon antigenic determinants present in mouse monoclonal antibodies. The raised anti-antibodies replaced [$I^125$ ]bGH bound to goat liver microsomes indicating production of anti-idiotypic antibodies against bovine growth hormone. These antibodies can have profound implications in vivo in lactating bovines for enhancing milk yield.

• Journal of Microbiology and Biotechnology
• /
• v.7 no.3
• /
• pp.189-193
• /
• 1997

Cryptosporidium parvum has been recognized as a significant cause of life-threatening diarrhea in Acquired ImmunoDeficiency Syndrome (AIDS) patients. Clinical diagnosis of cryptosporidial infections has been primarily based on the detection of infective stage, oocysts, in stools. Anti-Cryptosporidium oocyst monoclonal antibody (mAb), IgG2a, recognizing an antigen of 97 kDa was generated to be used for diagnosis of Cryptosporidium infection in AIDS patients using an immunofluorecence. It appeared to react with the surface antigens. Transmission electron micrographs of the infective stage of Cryptosporidium recognized by this mAb demonstrated sporolulated oocysts, which measure $4~6{\mu}m$, and sporozoites excysting from oocysts.

• Korean Journal of Veterinary Service
• /
• v.21 no.2
• /
• pp.127-132
• /
• 1998

To study the efficacy monoclonal antibody(MAb) against bovine rotavirus(BCV) in treatment of calf diarrhea, the MAb was fed to 166 calves with diarrhea from Chung-buk area. The results were summarized as follows. 1. Among the 1,049 calves investigated, 166(16%) calves were infected with BCV. 2. The monthly rate of BCV infection were higher in October to December compared with other months of the year. 3. Among the 166 calves with diarrhea, 137(83%) calves were recovered. 4. Young calves within 7-day-old were more effective in treatment than other ages and the rate of treatment was 86% 5. Most effective period for treatment of rotavirus was at the first stage of infection.