• Title/Summary/Keyword: 표면항체

Search Result 130, Processing Time 0.023 seconds

Ultrastructural Localization of a Common Antigen of Sporozoites and Merozoites of Cryptosporidium by Immunogold Labeling Technique Using a Monoclonal Antibody (Monoclonal Antibody와 Immunogold 표지법에 의한 Cryptosporidium의 Sporozoites와 Merozoites의 공통항원의 구조적 위치 결정)

  • Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
    • /
    • v.17 no.5
    • /
    • pp.499-503
    • /
    • 1989
  • Relatively little is known about the antigenic relatedness of the different developmental stages of Cryptosporidium. A monoclonal antibody (mAb), an IgG3, was produced against the Cryp-tosporidium merozoite stage by immunizing mice with merozoite preparation. This monoclonal was reacted with sporozoite antigens in Western blotting resulting in recognition of an epitope on a 3.5-kDa antigen. An immunoelectron microscopic technique was used to investigate the antigenic relatedness of Cryptosporidium Sporozoites and merozoites. Mouse intestine was fixed with 1 % glutaraldehyde and embedded in LR White. Thin sections were then sequentially treated with murine IgG3 mAb and anti-mouse IgG conjugated to 15-nm diameter colloidal gold. This mAb showed similar (sur-face/cytoplasmic) immunoelectron microsropic colloidal gold labeling patterns with sporozoites and merozoites, indicalting epitope sharing between these two stages. This information might be useful for identifying possible epitopes to which a vaccine could be developed.

  • PDF

Expression of Antibody Genes Specific for Human Hepatitis-B Virus in Transgenic Tabacco Plants (형질전환된 담배에서 사람 B형 간염바이러스 항체 유전자의 발현)

  • Seok Yoon KWON;Shin Je KIM;Hyo Jeong HONG;Moon Hi HAN;Chang Ho CHUNG;Ho Sul LEE;Kyung Hee PAEK
    • Korean Journal of Plant Tissue Culture
    • /
    • v.21 no.6
    • /
    • pp.353-356
    • /
    • 1994
  • Chimeric kappa chain and gamma chain cDNA clones (pCKS2 and pCHS2) of a monoclonal antibody specific for pre-S2 surface antigen of human hepatitis-B virus were ligated into Xbal site of plant expression vector pBKS-1. Plasmid DNA containing each of the chimeric gene were then mobilized from E, coli to Agrobacterium tumefaciens strain LBA4404. The chimeric antibody genes were then introduced into tobacco by Ti plasmid-mediated transformation. The putative Transformants were selected on medium containing kamaycin sulfate. Shoots that formed on shoot induction medium were analyzed by Western blot analysis for the expression of kappa-chain or gamma-chain genes. The Western blot analyses clearly showed that the introduced genes were stably expressed in transgenic plants.

  • PDF

Purification of Streptomyces Phospholipase D by Immunoaffinity Chromatoghraphy using Peptide Antibodies (Streptomyces phospholipase D의 정제를 위한 면역친화 크로마토그래피의 개발)

  • Park, In-Sun;Kim, Young-Ah;Jeong, Su-Jin;Uhm, Tai-Boong
    • Korean Journal of Microbiology
    • /
    • v.42 no.4
    • /
    • pp.294-298
    • /
    • 2006
  • An immunoaffinity chromatography for the specific binding of Streptomyces somaliensis phospholipase D (PLD) that is considered as an industrially potential enzyme was developed. By using the protein structure prediction programs and the X-ray crystal structure of a Streptomyces PLD, 5 different epitopes with high antigenicity that are predicted to locate on the surface of the S. somaliensis PLD were selected and then synthesized for the preparation of antipeptide antibodies. Each purified rabbit IgG was coupled with NHS-activated Sepharose to prepare the immunoaffinity resins. After one-step purification of the culture concentrate on the antipeptide IgG-coupled Sepharose column, SDS-PAGE and the Western blot analysis of the purified samples showed that purification of PLD on the affinity columns was satisfactory, indicating that the peptide design using the structural information of Streptomyces PLDs was rational. However, the purified PLD in the solution aggregated rapidly, which resulted in poor specific activity and low purification yield.

A research into perceptionality of students of the dental hygiene department on hepatitis B virus (치위생과 학생들의 B형 간염 바이러스에 대한 인지도 조사연구)

  • Kang, Eun-Ju
    • Journal of Korean society of Dental Hygiene
    • /
    • v.3 no.2
    • /
    • pp.89-99
    • /
    • 2003
  • This study aims to furnish basic data about prevention and infection control of Hepatitis B Virus(HBV) for those who, working in dental offices, are particularly exposed to a high risk of HBV infection. A survey was conducted to 310 students including freshmen, sophomores and juniors enrolled in the dental hygiene department in order to examine their knowledge about infection routes of HBV, clinical history of their family members and their own health. The outcomes of the survey showed following facts; 1. Students were found to lack knowledge about the present conditions of their HBsAg and HBsAb of HBV(PF0.05), conduct of preventive vaccination(PE0.05), completion of 3 required vaccinations(PF0.05) and formation of antibody(PF0.05). 2. Students named "blood"(88.6%) and "infected needles"(82.5%) as most likely infection routes of HBV(PE0.05 and PE0.01). These replies came mostly from sophomores(65.6% and 92.1%), followed by juniors(89.2%, 82.5%) and freshmen(81.1%, 73.0%). Least knowledge about infection routes of HBV was sensed with the reply "infection through breast-feeding of positive mother(27.9%)"(PE0.05). Generally, sophomores seemed to have much knowledge about infection routes, followed by juniors and freshmen in order. 3. As to clinical history of family members, 10 students(3.5%) replied that any of their family members is suffering from HBV now, 8(2.6%) revealed that some of their family members once suffered from it and 10(3.2%) reported cases of death of their family members from liver diseases. 4. Ninety-four point seven percent of respondents believed their health to be better than normal.

  • PDF

Integration of immunohistochemical reactions into Electrochemical and Optical Analyses of Biochips (면역 조직화학 반응이 통합된 바이오칩의 전기화학 및 광학적 분석)

  • Choi Hyoung Gil;Hong Eun Kyoung;Lee Seung-Won;Yoon Hyun C.
    • KSBB Journal
    • /
    • v.20 no.2 s.91
    • /
    • pp.123-128
    • /
    • 2005
  • We have addressed two important issues of immunosensing biochips, including the construction of antibody functionalized suface for efficient affinity reactions and the development of a signal registration strategy that converts biospecific reactions into highly quantifiable electrochemical and/or optical signals. The developed immunoassay reaction is an integrated version of enzyme-mediated immunoprecipitaion reaction, which is widely used in immunohistochemistry, and electrochemical signaling reaction. For the evaluation of analytical performance of fabricated immunosensing biochips, signaling for mouse IgG in antiserum was conducted. Applications of the developed strategy have been found for the evaluation of histology chemicals and for the signal amplification for array-type biochip analysis.

Fabrication of fiber-optic evanescent wave immunosensor and its measuring characteristics (광섬유 소산파를 이용한 면역 센서 제조 및 그 특성)

  • Choi, Ki-Bong;Youn, Hee-Ju;Cha, Seung-Hee;Choi, Jung-Do
    • Journal of Sensor Science and Technology
    • /
    • v.6 no.5
    • /
    • pp.356-361
    • /
    • 1997
  • Fiber-optic evanescent wave sensor was designed and fabricated to detect mouse immunoglobulin G(IgG) with decladed optical fiber on which anti-mouse IgG was immobilized. A sensitivity obtained by any direct or competitive method was lower than $1\;{\mu}g/m{\ell}$. Anti-mouse IgG was immobilized on 93.9% of core surface of optical fiber by simple adsorption method. The effect of postcoating using bovine serum albumin to remove non-specific binding was not observed. As the ratio of fluorescein to mouse IgG increased, the fluorescence signal increased, but that increase showed no linear relationship. Our fiber-optic sensor system could be used as immunosensor by measuring evanescent fluorescence in antigen-antibody reaction with good sensitivity below $1{\mu}g/m{\ell}$ level.

  • PDF

Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum (항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구)

  • Kim, S.C.;Lee, K.S.;Kim, Y.K.;Kim, C.K.;Choi, K.H.;Kwon, O.J.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.17 no.1
    • /
    • pp.71-80
    • /
    • 1990
  • New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

  • PDF

Recent Advances in Cancer Diagnosis: On an Overview of Diagnostic Cytopathologic Modalities and Ancillary Techniques (세포병리학적 기초에 의한 암진단의 발전: 진단방법과 보조기법)

  • Kim, Ki-Tai;Ham, Eui-Keun
    • The Korean Journal of Cytopathology
    • /
    • v.7 no.1
    • /
    • pp.1-11
    • /
    • 1996
  • From the concepts of cellular pathology and of exfoliative cytology, as elucidated by Virchow and Papanicolaou respectively in the late 19th and early 20th century, have evolved the primary methods for the diagnosis of cancer today. From Papanicolaou's concept of exfoliative cytology developed fine needle aspiration biopsy in the early 1960's, this has become a major diagnostic procedure and has contributed to a significant reduction in open biopsies and, therefore, to medical cost-effectiveness immunobiochemical techniques provided us with a supplement to cancer diagnosis in the 1980's. The immunoperoxidase method, using monoclonal antibodies, is applied primarily as an ancillary measure to elucidate the nature of cancers The availability of specific monoclonal antibodies has greatly facilitated the identification of cell products or surface markers. For example, antibodies directed against intermediate filaments have proved to be of value in determining the histogenesis oi poorly differentiated neoplasms. Tumor markers may serve as biochemical indicators of the presence of a neoplasm. They can be detected In plasma and other body fluids. Their concentration can be applied as a diagnostic test, for monitoring the clinical course of known cancer, and as a screening measure to detect certain cancers in a population at risk. Flow cytometry is a useful tool for distinguishing several cell characteristics, such as the immunophenotype of leukemia-lymphoma cells, the DNA content of neoplastic cells, and cell proliferation rate. Molecular biologic techniques provided a giant step for the management of cancer patients encompassing diagnosis, prognostic evaluation, and therapy. Nucleic acid hybridization techniques are utilized as Southern, Northern, and dot blots and in situ hybridization. Molecular biology and its techniques may bring a blight new horizon for understanding cancer biology and in designing therapy on the basis of gene manipulation.

  • PDF

Improvement of Artificial Antibody Secretion Using Supercharged Protein (단백질의 과전하화를 이용한 인공 항체의 분비 개선)

  • Park, Jiyeon;Choi, Heeju;Lee, Hyejin;Ahn, Jung Hoon
    • Journal of Life Science
    • /
    • v.30 no.5
    • /
    • pp.420-427
    • /
    • 2020
  • A repebody, an artificial non-immunoglobulin protein scaffold, is expected to be a solution in the search for faster, cheaper, and customizable antibodies. However, the production of medical repebodies remains difficult due to their low yield and the complex purification processes required. The Pseudomonas fluorescens ABC transporter system has been suggested as an efficient and cost-effective method for repebody production, but the total yield is low because of the secreted protein's positive charge; thus, a repebody with a high isoelectric point needs to be changed into a more negatively charged protein for better secretion. To achieve this, we first attached oligo-aspartic acids to the N- and C-terminals of the repebody, but secretion efficiency was not enhanced significantly. Subsequently, we devised an alternative method for improved secretion efficiency by engineering fifteen positively charged amino acids to aspartic acid in the non-antigen binding sites of the repebody to give a high net negative charge. As a result, secretion efficiency was greatly enhanced from 21.2% (wildtype) to 58.5% (negatively supercharged). The negatively supercharged repebody was succussfully produced extracellularly by ABC transporter secretion system in P. fluorescens.

cDNA microarray analysis of viral hemorrhagic septicemia infected olive flounder, Paralichthys olivaceus: immune gene expression at different water temperature (바이러스성 출혈성 패혈증에 감염된 넙치의 cDNA microarray 분석 : 수온에 따른 면역 유전자 발현의 차이)

  • Kim, Jin-Ung;Jung, Sung-Ju
    • Journal of fish pathology
    • /
    • v.27 no.1
    • /
    • pp.1-9
    • /
    • 2014
  • The olive flounder, Paralichthys olivaceus is susceptible to viral hemorrhagic septicaemia virus (VHSV) at $15^{\circ}C$ but no mortality at $20^{\circ}C$ even though the virus can grow well in vitro at $20^{\circ}C$. Thus, we designed an experiment to know immune response of olive flounder against VHSV when the host reared at $15^{\circ}C$ or $20^{\circ}C$. cDNA microarray analysis was performed to compare the gene expression patterns of the kidney cells between the host reared at $15^{\circ}C$ or $20^{\circ}C$. The expression of MHC class I, IL-8, myeloperoxidae and endonuclease G-like having function for the antigen presentation and chemokine-factor were up-regulted both the $15^{\circ}C$ and $20^{\circ}C$ during VHSV infection. MHC class II gene existing on antigen-presenting cells and B cell lymphocytes, immunoglobulin (Ig) genes and phagocytosis related genes were down-regulated at $15^{\circ}C$ but highly expressed at $20^{\circ}C$. It can be thought that innate immune related antigen presentation by MHC class I and phagocytosis reaction against VHSV are efficiently occur both the temperature but macrophage or B cell related antigen presentation via MHC class II fails to induce downstream immune reactions (adaptive immunity) to make antibody, and it can be one of the reason that causes high mortality only at $15^{\circ}C$.