• Journal of Microbiology
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• v.35 no.2
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• pp.87-91
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• 1997

Monoclonal antibodies were prepared in order to an assay method for Vibrio vulnificus. Sixteen mouse ybridoma cell lines were established by immunization of whole cell antigen to BALB/c mice, fusion with SP2/O myeloma cells, and cloning. Most of them secreted IgM.lambda. antibodies. A sandwich enzyme-linked immunosorbent assay was developed with rabbit anti-V. vulnificus polyclonal antibodies as capture antibody, an IgM monoclonal antibody as detector antibody, and goat anti-mouse IgM-alkaline phosphatase conjugate as developer antibody. The range of detection was 10$\^$4/ to 10 V. vulnificus cells per microplate well. When four related Vibrio species were tested for cross-reactions, V. parahaemolyticus showed 3.5% reactively and V. carchariae, V. fluvialis, and V. furnisii showed negligibal (<1%) cross-reactivity.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.269-272
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• 1989

In order to obtain monoclonal antibody from ascites fluid at sufficiently high purity using a single hydroxylapatite chromatography (HA) a further optimization on its operating variables was carried out. By adjusting the pH of the eluent, the sodium phosphate buffer, to 6.0 from 6.8 and adding CaCl$_2$to 1 mM at the column inlet, the elution molarities (M$_{elu}$) for the desired monoclonal antibody and contaminating proteins can be distinguished from each other with enough resolution. Previously these two groups of proteins co-eluted at the same time at pH 6.8 and without CaCl$_2$. This sin81e step hydroxylapatite chromatography yields the desired antibody pure enough for diagnostic use.

• YAKHAK HOEJI
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• v.34 no.1
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• pp.11-14
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• 1990

$NH_2-terminal$ amino acid sequence analysis of monoclonal antibody is very important to identify gene family and diversities of antigen-antibody recognition. When we used the PVDF (Polyvinylidene difluoride) membrane blotting method, we could easily analyze $NH_2-terminal$ sequence of monoclonal-antibody which specifically binds to phosphatidylinositol 4,5-biphosphate. PVDF membrane is an ideal solid-phase support for sequence analysis, especially when used with electroblotting method. This method is superior to continual method and will be applied to the sequence analysis of picomole quantities of proteins by gel electrophoresis.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.347-348
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• 1993

• Korean Journal of Veterinary Research
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• v.44 no.4
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• pp.625-635
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• 2004

Trichnuris suis does not excrete eggs during larval stage as well as in particular adult stage, It is impossible to diagnose by use of fecal examination method in those periods. Therefore, serological diagnostic method can be very useful for those stages. In order to produce monoclonal antibody, specific somatic and secretory-excretory (SE) antigens of T. suis were identified and analyzed by SDS-PAGE and Western blot. Monoclonal antibody-producing hybridoma cells were cloned, which were made of popliteal lymph node of BALB/c mice immunized with a 45 kDa somatic antigen of T. suis. Five clones (1B9, 2C4, n2C5, 2D7 and 2D8) showing strong responses to T. suis antigens were selected and the isotype identified. All monoclonal antibodies were IgG1 isotype and the light chains were k chain. Established monoclonal antibodies reacted specifically to somatic and SE antigens of T. suis and did not cross-reacted to antigens of ascaris suum, trichuris vulpis, or Trichinella spiralis. The sensitivity of somatic and SE antigens against these monoclonal antibodies were significant (p<0.01) associated with those of positive and negative sera.

• Nuclear Medicine and Molecular Imaging
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• v.40 no.2
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• pp.66-73
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• 2006

Monoclonal antibodies are designed to bind specifically to certain antigen, give therapeutic effect to the target and to be produced in large scale with homogeneity. The monoclonal antibodies conjugated with radionuclide can deliver therapeutic irradiation to the target, and showed successful results in certain malignancies, which is known as radioimmunotherapy. The target-to-background ratio depends on the antigen expression in the target and normal tissues, which is related to the therapeutic efficacy and toxicity in radioimmunotherapy. For the solid tumor beta-ray energy should be high, but lower beta energy is better for the hematological malignancies. I-l31 is widely used in thyroid cancer with low cost and high availability. Labeling monoclonal antibody with I-131 is relatively simple and reproducible. Some preclinical data for the I-131 labeled monoclonal antibodies including acute toxicity and efficacy are available from already published literatures in KIRAMS, physician sponsored clinical trial protocols using Rituximab, KFDA approved anti-CD20 chimeric monoclonal antibody and I-131 were approved by KFDA and currently are ongoing.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1498-1504
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• 2005

Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

• Biomolecules & Therapeutics
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• v.5 no.4
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• pp.344-350
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• 1997

The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.

• Korean Journal of Veterinary Research
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• v.30 no.4
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• pp.511-513
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• 1990

Monoclonal antibody to progesterone was produced using the antigen $11{\alpha}$-hydroxyprogesterone hemisuccinate conjugated to bovine serum albumin. Hybridomas secreting antibody to progesterone were detected by radioimmunoassay and enzyme-linked immunosorbent assay and cloned in soft agar. Two stable monoclonal antibodies which were highly specific to progesterone were obtained, so it may be advantageously used to study on several physiological functions of progesterone including immunological research.

• Journal of Microbiology
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• v.34 no.4
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• pp.379-383
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• 1996

Cryptosporidium parvum causes a life-threatening diarrhea in acquired immunodeficiency syndrome (AIDS) patients. THe sporozoite stage of C. parvum has been known to be a target in treating cryptosporidiosis in AIDS patients as it is an extracellular stage. A sporozoite was ultrastructurally observed. It has a creascent shape with a rounded posterior end and a tapering body. The compact nucleus was located at the posterior end. A monoclonal antibody was produced, which recognized a 43 kDa of sporozoite antigens in a western blot analysis and showed the surface labeling in immunofluorescence.