• Bulletin of the Korean Chemical Society
• /
• v.23 no.8
• /
• pp.1116-1119
• /
• 2002

Monoclonal antibodies have been generated against a generic hapten, ο,ο-diethyl ο-(5-carboxy-2-fluorophenyl) phosphorothioate, for the determination of organophosphorus (OP) pesticides in a class-specific manner. In an indirect competitive enzyme-linked immunosorbent assay (ELISA) format, employing a heterologous coating antigen, these monoclonal antibodies showed desirable properties for use in the class-specific determination, i.e., broad specificity and high sensitivity. The IC50 values of four commonly used ο,ο-diethyl OP pesticides were fairly uniform ranging from 0.1 to 0.3 ㎛/mL. The IC50 values of three ο,ο-dimethyl derivatives were between 0.3 and 1.4 ㎛/mL. These values, together with the limits of detection (LOD), were better, in terms of the specificity and sensitivity, compared with the values obtained previously with polyclonal antibodies.

• Archives of Pharmacal Research
• /
• v.22 no.3
• /
• pp.262-266
• /
• 1999

Dopamine transporter (DAT) plays the most important role in terminating the actions of dopamines released into the synaptic cleft. DAT is also the target of various psychotropic drugs such as cocaine and amphetamine. In this study were prepared DAT-specific antibodies using the 2nd extracellular loop of rat DAT as an antigen. The 2nd extracellular loop of the rat DAT was expressed in bacterial as a fusion protein with glutathione-S-transferase, and injected ito rabbits to raise antibodies. Produced antibodies clearly recognized the rat DAT in ELISA, immunoblotting, and immumoprecipitation. As expected from the high sequence homology between the rat and human DAT, the antibodies raised for the rat DAT cross-reacted with the human DAT in the immunoblotting. Considering the specificity for DAT with wide range of applications such as ELISA, immunoblotting, and immunoprecipitation, these antibodies would be valuable tool for understanding the pharmacological actions of dopamine transporter and drug addition.

• Parasites, Hosts and Diseases
• /
• v.37 no.4
• /
• pp.243-248
• /
• 1999

The present study analyzed serum IgG subclass antibody reaction to major antigenic bands of Clonorchis sinensis to investigate improvement of its serodiagnosis. Of the four subclass antibodies, IgG1 and IgG2 antibodies were produced but not specific, IgG3 antibody was least produced, and IgG4 antibody was prominent and specific. The serum IgG antibody reaction to any of 43-50, 34-37, 26-28, and 8 kDa bands was found in 65.5％ of 168 egg positive cases while IgG4 antibody reaction was found in 22.0％ of them. The positive rates of IgG and IgG4 antibodies were directly correlated with the intensity of infection. All of the sera from heavily infected cases over EPG 5,000 showed positive reaction for specific IgG and IgG4 antibodies. The specific serum IgG4 antibody disappeared within 6 months after treatment. The bands of 35 kDa and 67 kDa cross-reacted with IgG antibodies but not with IgG4 antibodies in sera of other trematode infections. The present findings suggest that serum IgG4 antibody reaction to 8 kDa band is specific but not sensitive. Any method to increase its sensitivity is required for improved serodiagnosis.

• 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.

• IMMUNE NETWORK
• /
• v.1 no.1
• /
• pp.7-13
• /
• 2001

Currently 18 monoclonal antibodies were approved by FDA for inj ection into humans for therapeutic or diagnostic purpose. And 146 clinical trials are under way to evaluate the efficacy of monoclonal antibodies as anti-cancer agents, which comprise 9 % of clinical trials in cancer therapy field. When considering a lot of disappointment and worries existed in this field during the past 15 years, this boom could be called as resurrection. Antibodies have several merits over small molecule drug. First of all it is easier and faster in development, as proper immunization of the target proteins usually raises good antibody response. The side effects of antibodies are more likely to be checked out in immunohistomchemical staining of whole human tissues. Antibody has better pharmacokinetics, which means a longer half-life. And it is non-toxic as it is purely a "natural drug. Vast array of methods was developed to get the recombinant antibodies to be used as drug. The mice with human immunoglobulin genes were generated. Fully human antibodies can be developed in fast and easy way from these mice through immunization. These mice could make even human monoclonal antibodies against any human antigen like albumin. The concept of combinatorial library was also actively adopted for this purpose. Specific antibodies can be screened out from phage, mRNA, ribosomal library displaying recombinant antibodies like single chain Fvs or Fabs. Then the coding genes of these specific antibodies are obtained from the selected protein-gene units, and used for industrial scale production. Both $na\ddot{i}ve$ and immunized libraries are proved to be effective for this purpose. In post-map arena, antibodies are receiving another spotlight as molecular probes against numerous targets screened out from functional genomics or proteomics. Actually many of these antibodies used for this purpose are already human ones. Through alliance of these two actively growing research areas, antibody would play a central role in target discovery and drug development.

• Parasites, Hosts and Diseases
• /
• v.51 no.6
• /
• pp.755-757
• /
• 2013

Gnathostoma spinigerum can cause subarachnoid hemorrhage (SAH). The detection of specific antibodies in serum against G. spinigerum antigen is helpful for diagnosis of neurognathostomiasis. There is limited data on the frequency of G. spinigerum infection in non-traumatic SAH. A series of patients diagnosed as non-traumatic SAH at the Srinagarind Hospital, Khon Kaen University, Thailand between January 2011 and January 2013 were studied. CT or MR imaging of the brain was used for diagnosis of SAH. Patients were categorized as aneurysmal subarachnoid hemorrhage (A-SAH) or non-aneurysmal subarachnoid hemorrhage (NA-SAH) according to the results of cerebral angiograms. The presence of specific antibodies in serum against 21- or 24-kDa G. spinigerum antigen was determined using the immunoblot technique. The detection rate of antibodies was compared between the 2 groups. Of the 118 non-traumatic SAH patients for whom cerebral angiogram and immunoblot data were available, 80 (67.8%) patients had A-SAH, whereas 38 (32.2%) had NASAH. Overall, 23.7% were positive for specific antibodies against 21- and /or 24-kDa G. spinigerum antigen. No significant differences were found in the positive rate of specific antibodies against G. spinigerum in both groups (P-value=0.350).

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.1
• /
• pp.17-24
• /
• 1998

We have previously demonstrated that specific antigens involved in autoimmunity in endometriosis may be endometrial proteins with molecular weight (mw) of 71, 92, and 103 kilodalton (kDa). The purposes of this study were to determine the incidence of IgG antibodies against these endometrial antigens in peritoneal fluid of patients with endometriosis and to evaluate the antigenic differences between the endometria of patients with and without endometriosis. Forty peritoneal fluid (PF) from 24 patients with endometriosis and 16 patients without endometriosis (control patients) were tested against endometrial protein from patients (n=8) with endometriosis and from control patients (n=10) by western blot. Fifteen (62.5%) of 24 PF samples from patients with endometriosis had specific Immunoglobuiin (Ig) G antibodies against one of three endometrial proteins with mw of 71, 92 and 103 kDa but none of PF samples from control patients had these antibodies. The electrophoretic pattern of endometrial proteins from patients with endometriosis was similiar to that from control patients. Furthemore there was no significant difference in specific PF Immunoglobulin G binding to endometrial proteins regardless of origin of these proteins. Our data indicate that specific humoral immune response can be found in PF of patients with endometriosis and that specific antigens inducing this immune response are present in human endometrium and that there is no antigenic difference between the endometria of patients with and without endometriosis.

• IMMUNE NETWORK
• /
• v.9 no.5
• /
• pp.203-207
• /
• 2009

Background: The FimA of Porphyromonas gingivalis is a crucial pathogenic component of the bacteria and has been implicated as a target for vaccine development against the periodontal diseases. Methods: In this study, the purified fimbriae (FimA subunit polymers) protein was used for immunization in their native form and B hybridoma clones producing antibodies specific to FimA were established. Results: The monoclonal antibodies prepared from selected two clones, designated #123 (IgG2b/ kappa) and #265 (IgG1/kappa), displayed different patterns of binding activity against the cognate antigen. Both antibodies reacted with conformational epitopes expressed by partially dissociated oligomers, but not with monomer as elucidated by Western blot analysis. Ascites fluid containing the monoclonal antibodies showed the inhibitory activity against P. gingivalis to saliva-coated hydroxyapatite beads, an in vitro model for the pellicle-coated tooth surface. Conclusion: These results suggest that the monoclonal antibodies could be used as vaccine material against the periodontal diseases through passive immunization.

• Parasites, Hosts and Diseases
• /
• v.29 no.3
• /
• pp.293-310
• /
• 1991

Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

• Journal of Life Science
• /
• v.5 no.2
• /
• pp.63-69
• /
• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.