• BMB Reports
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• 제34권1호
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• pp.33-38
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• 2001

The human PTK6 (also known as Brk) polypeptide, which is deduced from its full-length cDNA, represents a non-receptor protein tyrosine kinase (PTK). It contains SH3, SH2, and tyrosine kinase catalytic domains that are closely related to Src family members. We generated an antihuman PTK6 antibody by immunizing rabbits with a PTK6-specific oligopeptide conjugated to BSA, which corresponds to 11 amino acid residues near the C-terminus. An immunoblot analysis with the antibody detected an expected 52-kDa band in various mammalian transformed cell lines. Immunoprecipitation and immunoblot analyses demonstrated that PTK6 is phosphorylated on the tyrosine residues) and interacts with approximately a 23-kDa tyrosine-phosphorylated polypeptide (most likely a substrate of PTK6) in breast carcinoma T-47D cells. An immunofluorescence analysis demonstrated that PTK6 is localized throughout the cytoplasm of T-47D cells. These results support a possible role for PTK6 in the intracellular signal transduction through tyrosine phosphorylation.

• Nutritional Sciences
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• 제7권1호
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• pp.31-34
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• 2004

This paper reviews the effects of Spirulina platensis and its extracts and phycocyanin, a blue photosynthetic pigment protein in Spirulina on the mucosal immune functions in humans and animals as follows: TEX>$\bullet$ IgA antibody response and other classes in mucosal immunity of mice treated with Spirulina platensis and its extract. $\bullet$ Effect of Spirulina phycocyanin ingestion on the mucosal antibody responses in mice. - Distinct effects of phycocyanin on secretory IgA and allergic IgE antibody responses in mice following oral immunization with antigen-entrapped biodegradable microparticles. $\bullet$ Influence of dietary Spirulina platensis on IgA level in human saliva. $\bullet$ A study on enhancement of bone-marrow cell-proliferation and differentiation by Spirulina platensis in mice: in vivo and in vitro study

• 대한미생물학회지
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• 제14권1호
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• pp.63-69
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• 1979

Antibodies to Propionibacterium acnes(Corynebacterium parvum) serotype I and serotype II in normal human sera were measured using a microtitre bacterial agglutination test. Of 168 sera tested, 53 sera(31.0%) exhibited higher agglutinin titres to serotype I than to serotype II and 34 sera(20.2%) gave higher titers to serotype II than to serotype I. Eighty-one sera(48.3%), however, showed similar antibody titres to both types. Antibodies to serotype I(x) and serotype II(y) showed high correlation(r=0.73, p<0.01) and regression equation was Y=1,078+0.73X. The mean antibody titre($log_2$) of 529 normal sera(male 447 and female 82) to serotype I was $5.49{\pm}1.36$, but there was no significant difference between male($5.45{\pm}1.36$) and female($5.74{\pm}1.36$). Bacterial agglutinin to Propionibacterium acnes in normal sera belonged to a 2-mercaptoethanol resistant IgG class.

• Molecules and Cells
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• 제27권2호
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• pp.225-235
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• 2009

Antibody phage display provides a powerful and efficient tool for the discovery and development of monoclonal antibodies for therapeutic and other applications. Antibody clones from synthetic libraries with optimized design features have several distinct advantages that include high stability, high levels of expression, and ease of downstream optimization and engineering. In this study, a fully synthetic human scFv library with six diversified CDRs was constructed by polymerase chain reaction assembly of overlapping oligonucleotides. In order to maximize the functional diversity of the library, a ${\beta}$-lactamase selection strategy was employed in which the assembled scFv gene repertoire was fused to the 5'-end of the ${\beta}$-lactamase gene, and in-frame scFv clones were enriched by carbenicillin selection. A final library with an estimated total diversity of $7.6{\times}10^9$, greater than 70% functional diversity, and diversification of all six CDRs was obtained after insertion of fully randomized CDR-H3 sequences into this proofread repertoire. The performance of the library was validated using a number of target antigens, against which multiple unique scFv sequences with dissociation constants in the nanomolar range were isolated.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.595-598
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• 2000

Aeromonas hydrophila is a pathogen to fish as well as human. It is a food-borne disease, and causes severe mortality in fish, and sometimes severe septicemia in human. In this study, a rapid detection method using latex agglutination has been developed for A. hydrophila. Polyclonal antibodies were raised against membrane and whole cells of three isolates from rainbow trout. Among these, latex particles coated with antibodies raised against whole cells of isolate No. 2 showed the best sensitivity. With latex particles coated with this antibody, we could detect $5{\times}10^4$ CFU of A. hydrophila in 5 min. The cross-reactivity with bacteria constituting the normal intestinal microflora and other pathogens for rainbow trout was insignificant. This latex agglutination assay method produced positive reaction with all clinical isolates of A. hydrophila which were identified by species-specific PCR for 16S rRNA in A. hydrophila.

• Bulletin of the Korean Chemical Society
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• 제28권10호
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• pp.1811-1814
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• 2007

• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.246-256
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• 1983

폐흡충증의 유행정도를 측정하는 데 사용하여 온 피내반응과 충란검사가 각각 특이도와 민감도에 문제를 갖고 있어 주민 중 현증 환자를 적절히 찾았다고 하기가 어렵게 되어가고 있다. 이러한 점을 개선하는 방법으로 폐흡충 특이 IgG항체를 검사함으로써 항체양성자의 수를 현증 환자의 수로 하는 방법을 폐흡충증 역학조사에 응용하고자 하였다. 이 연구에서는 특이항체를 면역효소진단법으로 측정하였을 때 그 효용성을 평가하고자 하였다. 전라남도 해남군 북평면과 북일면의 6개 국민학교, 2개 중학교, 1개 고등학교 학생과 2개 부락 주민 등 모두 4,285명을 우선 피내 반응으로 검사하고 폐흡충 피내반응 양성자에서 가래 검사를 한번 실시하였고 혈청을 채취하여 면역효소진단법으로 폐흡충 특이 IgG항체를 측정하였다. 그 결과를 요약하면 다음과 같다. 1. 총 대상자 4,285명 중 폐홉충 피내반응 양성자는 244명 (5.7%)이었고, 피내반응 양성자 중 충란 양성자는 7명 (4.2%), 면역효소진단법에 의한 특이 IgG항체 양성자는 40명 (16.7%)이었다. 전체 조사 대상자에 대한 충란 양성자의 비율은 0. 16%, 특이 IgG항체 양성자는 0.93%에 해당하였다. 2. 폐흡충 특이 IgG항체 양성자는 충란 양성자에서는 100%, 폐흡충 피내 반응은 양성이나 충란검사 음성자에서는 14.2%에서 나타났으나, 간흡충 피내반응 양성자 또는 피내반응 음성 자에서는 나타나지 않았다. 3. 폐흡충 특이 IgG항체 양성자는 피내반응 구진의 크기 9mrn이하에서는 없었고, 10mm에서는 5.5%, 11∼12mm에서는 26.1% 및 18.2%, 13mm이상에서는 평균 42.4%에서 양성이었다. 4. 특이항체 양성자 31명을 임상적으로 조사한 바 검사소건(흉부 X-선, 충란검사 및 호산구 비율)에서 폐흡충증에 합치하는 이상소견이 24례에서 나타나 현증 폐홉충증 환자로 생각할 수 있었고, 나머지 7례중 3례는 폐흡충증의 과거력이나 쳔증 증상을 갖고 있었으며 4례에서는 특이항체 양성소견 이외에는 어느 검사로도 폐흡충증임을 증명할 수가 없었다. 5. 폐홉충 특이 IgG항체 양성자 수의 피내반응 양성자 수에 대한 비율은 연령별로 7∼9세에서는 16.6%, 10∼14세에서 25.4%, 15w18세 군에서는 18.3%이었고 20세 이상에서는 10.0%로 떨어지고 있었다. 6. 체홉충 특이 IgG항체 양성자는 북평면의 산골 마을에서 주로 나타나고 있었다. 이상과 같은 결과에서 면역효소 진단법에 의하여 폐흡충 특이 IgG항체 양성자를 조사함으로써 어느 지역의 폐흡충중 현증 환자를 피내 반응이나 충란 검사보다도 더 사실에 가깝게 측정할 수 있다고 생각하였다. (이 연구를 수행하는데 현지조사를 직접 간접으로 도와 주신 해남군 보건소 민 경 소장님과 김용팔 주사, 북평면 면사무소 직원여러분과 해남군 교육청, 각급학교 교직원 여러분께 감사드립니다. 환자의 흉부 X-선 사진을 판독하여 주신 중앙의대 부속 성심병원 방사선과 과장 정전상 부교수께 도 감사드립니다. )

• 대한수의학회지
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• 제37권3호
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• pp.547-554
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• 1997

Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

• Toxicological Research
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• 제14권2호
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• pp.123-127
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• 1998

BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

• Journal of Microbiology and Biotechnology
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• 제16권7호
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• pp.1104-1110
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• 2006

Recombinant antibody-toxin is a bifunctional protein that binds and kills a target cell expressing a specific antigen on the surface of the cell, and its structure is chimeric, in which a toxin is fused to an antigen-binding domain such as scFv or Fab. Divalent antibody-toxin molecules showed higher cytotoxicities against cancer cell lines than monovalent molecules. However, the yields of the divalent molecules were very low. In this study, we introduced the CH2, CH3, or CH2-CH3 (=Fc) domain of antibody in the middle of the Fab-toxin between the hinge region of human IgG1 and the toxin domain to increase the yield. The covalently bonded dimer could be formed by three disulfide bridges from cysteine residues in the hinge region. The molecule with the CH3 domain showed about 3-fold higher dimerization yield than previously constructed Fab-toxin molecules, while maintaining the cytotoxic activity comparable to that of scFv-toxin. However, the introduction of CH2 or Fc domain to the same position showed little effect on the dimerization yield. We also observed that the introduction of the CH3 region made it possible to form noncovalently associated dimer molecules.