• Journal of Microbiology and Biotechnology
• /
• v.16 no.8
• /
• pp.1316-1319
• /
• 2006

An antibody displayed phage collection (SBAE-2R), screened from a human synthetic phage antibody library (Fab 21ox), was used for the determination of dextran. The dextran-binding affinity was determined by serologically specific transmission electron microscopy (TEM) and a paper dipstick assay. The phage collection was distributed over the dextrancoated grids with 39$\pm$25 phages/$\mu$m$^2$ on the grids. Phages were not seen on dextran-coated grids exposed to the Fab 2lox phage library. The phage collection (SBAE-2R) produced 54$\pm$3 color normalized intensity (N.I.) from 125 ppm to 1,000 ppm of dextran and 5$\pm$1 (N.I.) for 63 ppm of dextran in a paper dipstick assay. This research extends the analytical options for dextran analysis by antibody displayed phage with a minimum of equipment usage.

• Food Science of Animal Resources
• /
• v.18 no.2
• /
• pp.149-156
• /
• 1998

Chicken beef pork meats and isolated soy protein (ISP) were heated at 10$0^{\circ}C$ for 30min and then heat-resistant proteins were fractionated to examine cross-resistant protein from chicken meat but not with beef pork or ISP. Dot blotting using the polyclonal antibody showed that the sen-sitivity for detecting chicken meat was 1$\mu$m and antibody-antigen reaction was dose-dependant. Results of dot blotting analysis to compare the amount of chicken meat present in arket meat products(Kentucky Frank sausage;chicken meat 46.52% and pork 24.92% vs Bulgogi Ham;chicken meat 28.89% and turkey 31.44%)showed that the significant differences between two meat products in terms of chicken meat concentrations. Dose-dependant dot-blotting reaction was also observed in chicken meat samples with various dilution.

• Korean Journal of Veterinary Service
• /
• v.23 no.2
• /
• pp.113-124
• /
• 2000

A total of 500 sera and lungs of slaughtered pigs were examined to investigate antibody titer, prevalence of pulmonary lesion, and interrelation among lung lesion score, type of pulmonary lesion and antibody titers by ELISA. The results obtained were as follows ; 1. The highest distribution of antibody titer was showed at 20 - 80 in M hyopneumoniae, 160-640 in P multocida type A and 160 - 640 in A pleuropneumoniae serotype 2 and 5. 2. The prevalence of pulmonary lesions was 84.0%, mean pulmonary lesion and mean lung score listed as 24.0$\pm$19.8% and 2.5$\pm$1.6, respectively. 3. In the prevalence of type of pulmonary lesion, enzootic pneumonia, pleuropneumonia and pleuritis were 58.2%, 10.0% and 15.8%, respectively. 4. Lung lesion score and type of pulmonary lesion were not interrelated with the distribution of antibody titer to specific pathogens, and causative pathogens of respiratory diseases were complicated with various bacteria.

• Journal of the korean veterinary medical association
• /
• v.14 no.3
• /
• pp.189-191
• /
• 1978

• KSBB Journal
• /
• v.17 no.1
• /
• pp.20-25
• /
• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• Parasites, Hosts and Diseases
• /
• v.21 no.2
• /
• pp.246-256
• /
• 1983

As epidemiological parameters of human paragonimiasis, the positive rates of intradermal test and the sputum/stool ekaminations have long been employed in population surveys. However, both the specificity of the intradermal test and the sensitivity of sputumjstool examination have been gradually declined as the endemicity was lowered; thus the gap between above two parameters widened. In such context, the development of a new epidemiologic parameter or tool which makes it possible to accurately discriminate the active paragonimiasis cases was necessary. In the present study, the detection rate of Paragonimus-speclac IgG antibody by micro-ELISA was evaluated as an indicator of epidemiologic status of human paragonimiasis in a population. A total of 4, 285 students and inhabitants living in Bukpyeong Myeon and Bukil Myeon, Haenam Gun, Jeonlanam Do was surveyed in October, 1983 by intradermal test first. Out of them, 244 cases (5.7%) were found positively reacted to VBS antigen of F. westermani. Out of 168 positive reactors, 7 cases (4.2%) were egg positive either by two times of sputum examination or by one stool examination. That indicated that only 0. 16% of total surveyed were confirmed as active paragonimiasis by egg detection. When sera collected from 239 positive reactors of Intradermal test were tested by micro-ELISA for their specific IgG antibody, 40 cases(16.7%) were found to be positive. All of 7 egg positive cases were again positive for specific IgG antibody. Among remaining 232 intradermal test positive cases, 33 cases were positive for IgG antibody. In contrast to those, none of 42 positive reactors to intradermal test for Cloncrchis and of 128 intradermal test negative cases showed positive for Paragcnimus-specIfic IgG antibody. The rate of specific IgG antibody as detected by micro-ELISA appeared to be increased with the wheal size of the intradermal test. When the wheal sixte was over 13mm in diameter, about 50% of them were positive for specific IgG antibody. Thirty-one specific antibody positive cases were clinically evaluated by laboratory examinations (repeated sputum examination, peripheral eosinophil count and chest roentgenogram) and by history taking. Out of them 24 cases were associated with one or more positive laboratory findings: thus considered as active paragonimiasis cases. Out of 7 lab. finding-free cases 3 revealed past history of typical paragonimiasis symptoms, suggesting that they were in chronic or in convalescent stages. The remaining 4 cases were considered as either mild or ectopic infection cases; the possibility of cross-reaction with other helminthiases could not be ruled out. From the above results, it was inferred that the detection of Paragonimus-specIfic IgG antibody by micro-ELISA was very much helpful in detecting the active cases as well as in proper evaluation of the endemicity of human paragonimiasis in a population. The convenience of mass haildling of sera in micro-ELISA was considered another superiority as an epidemiologic tool.

• IMMUNE NETWORK
• /
• v.3 no.1
• /
• pp.16-22
• /
• 2003

Background: The S100A2 gene, also known as S100L or CaN19, encodes a protein comprised of 99-amino acids, is a member of the calcium-binding proteins of EF-hand family. According to a recent study, this gene was over-expressed in several early and malignant carcinomas compared to normal tissues. To elucidate the role of S100A2 protein in the process during carcinogenesis, production of monoclonal antibody specific to the protein is essential. Methods: First, cDNA sequence coding for ORF region of human S100A2 gene was amplified and cloned into an expression vector to produce GST fusion protein. Recombinant S100A2 protein and subsequently, monoclonal antibody to the protein were produced. The specificity of anti-S100A2 monoclonal antibody was confirmed by immunoblot analysis of cross reactivity to other recombinant proteins of S100A family (GST-S100A1, GST-S100A4 and GST-S100A6). To confirm the relation of S100A2 to cervical carcinogenesis, S100A2 protein in early cervical carcinoma tissue was immunostained using the monoclonal antibody. Results: GST-S100A2 recombinant protein was purified by affinity chromatography and then fusion protein was cleaved and S100A2 protein was isolated. The monoclonal antibody (KK0723; Korean patent pending #2001-30294) to the protein was produced and the antibody did not react with other members of EF-hand family proteins such as S100A1, S100A4 and S100A6. Conclusion: These data suggest that anti-S100A2 monoclonal antibody produced in this study can be very useful for the early detection of cervical carcinoma and elucidation of mechanism during the early cervical carcinogenesis.

• Parasites, Hosts and Diseases
• /
• v.29 no.4
• /
• pp.389-396
• /
• 1991

Antibody changes in experimental anisakiasis were observed in 10 rabbits which were infected each with 10 Anisakis simplex larvae. The sera were collected before and on the 6th to the 95th day after the infection. Using crude saline extract of Anisakis larvae as antigen, specific IgM and IgG antibody levels were observed by ELISA and SDS-polyacrylamide gel electrophoresislimmunoblot. Levels of specific-IgM antibody were elevated from the 6th day, reached their peaks on the lIth day after the infection, and dropped thereafter. Serum levels of IgG antibody increased from the 6th day and reached their peak on the 26th day after the infection, and decreased gradually thereafter. When SDS-PAGE of the crude extract was done, at least forty-one SDS-polypeptide bands were recognized. Of them, IgM antibody reacted mainly to the bands of 168, 95, 74, 64, 51, 47 and 34 kDa while IgG antibody reacted strongly to 168, 92, 85, 64, 58, 52, 42 and 40 kDa bands. The crude extract showed negligible cross reactions with sera of other parasitic diseases and normal control. Key words: Anisakis simplex larvae, experimental anisakiasis, rabbit, antibody, ELISA, SDS- PAGE/immunoblot.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.10 no.4
• /
• pp.309-314
• /
• 2005

We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.