• Journal of Plant Biology
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• 제39권2호
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• pp.99-105
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• 1996

Immunocytochemistry utilizes the specificity of the antigen-antibody reaction to localize specific antigens in cells or cellular organelles. Here we report the use of monoclonal antibodies, in conjunction with gold-labeled second antibodies to study the ultrastructural localization and tissue distribution of the Mr 98, 000 anionic peroxidase and other wall antigens. The antibody specific for this wall peroxidase, mWP3, labeled mainly the cell wall area. At the tissue level, the Mr 98, 000 peroxidase is located predominantly in the leaf mesophyll, internal coleoptile and sieve elements, but not in the root, as assayed with these procedures. The coleoptile walls were less heavily stained than the walls of leaf mesophyll cells. At the subcellular level, it is localized mainly in intercellular regions of the cell walls. A similar staining pattern was revealed by mWP19, one of anti-$\beta$ glucosidase antibody, though it looked less heavily stained than one with mWP3. In order to serve as a control wall staining using IgM monoclonal antibodies, mWP18 was used. Most of the label is localized over wall regions of cells of the young leaf mesophyll and coleoptile.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4255-4261
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• 2012

The present study was conducted to evaluate radioimmunoimaging (RII) and in vivo distribution of mixed antibodies $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb in nude mice bearing human lung adenocarcinoma xenografts. Single and mixed applications of the two radiolabeled monoclonal antibodies (mAbs) were compared. Direct labeling of $^{99m}Tc$ was applied to radiolabel the EGFR and CD44 mAbs. The properties of the radiolabeled antibodies were then characterized. RII and assessment of the distribution of the antibodies in nude mice bearing lung adenocarcinoma xenografts were achieved by applying separate and combined doses of $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb. The labeling rates of $^{99m}Tc$ for EGFR-mAb and CD44-mAb were $91.5%{\pm}3.8%$ and $92.3%{\pm}4.1%$ respectively, with specific activities of 2.8 and $2.9MBq/{\mu}g$, respectively, and radiochemical purities (RCP) of 96.5% and 96.2%. The radioactivity uptake of the combined application of both radiolabeled antibodies was clearly higher than with a single application of either alone. The relative values of target-to-nontarget (T/NT) measured through the regional interest (ROI) technique were $5.59{\pm}0.42$ (mixed antibodies), $2.78{\pm}0.20$ ($^{99m}Tc$-EGFR-mAb), and $2.28{\pm}0.16$ ($^{99m}Tc$-CD44-mAb) in the RII. The body distribution of the radiolabeled antibodies and their imaging results were basically identical. Application of the mixed antibodies with $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb can increase the radioactivity uptake of tumor tissue, leading to more ideal target-to-nontarget ratios, and therefore superior results.

• BMB Reports
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• 제37권5호
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• pp.556-564
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• 2004

A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2% glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

• 환경위생공학
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• 제4권1호
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• pp.17-28
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• 1989

Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

• Journal of Yeungnam Medical Science
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• 제7권2호
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• pp.27-37
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• 1990

흰 쥐 내피세포의 배양에 있어서 세포골격단백의 변화 양상과 심근세포와의 공존 배양시의 영향 그리고 colchicine 이 미치는 영향에 대해 알아보고자 monoclonal antibodies를 이용한 간접면역형광법으로 actin과 tubulin염색을 시행하여 microtubule과 microfilament의 변화 양상을 일령별로 관찰하였다. 실험결과를 요약하면 다음과 같다. 세포질이 넓게 퍼지고 많은 돌기를 가진 세포에서 섬유사형태의 반응이 강하였으며 내피세포가 단층을 형성하면서 섬유사형태의 반응이 약해졌다. 심근세포를 혼합배양한 군에서 내피세포가 단층을 형성하면서 microtubule은 대조군과 같은 수준으로 섬유사형태의 반응이 감소하였으나 microfilament는 여전히 강한 섬유사형태의 반응을 유지하였다. Colchicine을 처리한 군에서 microfilament는 대조군과의 차이점이 발견되지 않았으나 microtubule에서는 colchicine 처리 직후부터 섬유사 형태의 감소경향을 보였으며 1일 이후가 가장 현저한 감소를 보이다가 2일 이후에는 대조군의 수준으로 돌아왔다.

• Fisheries and Aquatic Sciences
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• 제8권4호
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• pp.213-219
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• 2005

Vitellogenin (VTG) was purified from the blood plasma of estradiol-17$\beta$ ($E_2$)-treated male marbled sole (Limanda yokohamae) using gel filtration and anion exchange chromatography. The purity of the marbled sole VTG (msVTG) was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequencing. The purified msVTG was used to produce monoclonal and polyclonal antibodies in mice and rabbits, respectively, and the specificity of the polyclonal antisera for msVTG was confirmed by Western blot analysis. The antibodies cross­reacted with a protein of molecular mass approximately 160 kDa in the plasma samples of mature female marbled sole. No cross-reactivity was observed with the plasma of male fish. A direct non-competitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed using the monoclonal anti-msVTG and polyclonal anti-msVTG antibodies, with purified msVTG as the standard protein. The values of the intra- and inter-assay variations were within the ranges of $8.l-9.8\%$ and $8.5-12.2\%$, respectively. The sensitivity was about 0.3 ng/mL. Serial dilutions of plasma from mature female sole reacted with the msVTG-antibodies in the sandwich ELISA, whereas the plasma from male fish did not. The results indicate that the maturation status of female marbled sole can be identified using a sandwich ELISA for msVTG.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.713-714
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• 2000

Six monoclonal antibodies to human cardiac troponin I (hcTnI) were produced to eventually develop an immunosensor for acute myocardial infarction (AMI). For the characterization of these antibodies, a set of 11 different peptides covering selected ranges of the complete amino acid sequence of hcTnI was prepared and used for epitope mapping. Such analysis allowed to select an appropriate pair of antibodies that can form a sandwich type of immune complexes and was consequently used for an immunoassay.

• 한국축산식품학회지
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• 제40권1호
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• pp.132-144
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• 2020

The aim of this study was to optimize staining procedures for muscle fiber typing efficiently and rapidly in bovine and porcine skeletal muscles, such as longissimus thoracis, psoas major, semimembranosus, and semitendinosus muscles. The commercially available monoclonal anti-myosin heavy chain (MHC) antibodies and fluorescent dye-conjugated secondary antibodies were applied to immunofluorescence histology. Two different procedures, such as cocktail and serial staining, were adopted to immunofluorescence analysis. In bovine muscles, three pure types (I, IIA, and IIX) and one hybrid type, IIA+IIX, were identified by the cocktail procedure with a combination of BA-F8, SC-71, BF-35, and 6H1 anti-MHC antibodies. Porcine muscle fibers were typed into four pure types (I, IIA, IIX, and IIB) and two hybrid types (IIA+IIX and IIX+IIB) by a serial procedure with a combination of BA-F8, SC-71, BF-35, and BF-F3. Unlike for bovine muscle, the cocktail procedure was not recommended in porcine muscle fiber typing because of the abnormal reactivity of SC-71 antibody under cocktail procedure. Within the four antibodies, combinations of two or more anti-MHC antibodies allowed us to distinguish pure fiber types or all fiber types including hybrid types. Application of other secondary antibodies conjugated with different fluorescent dyes allowed us to get improved image resolution that clearly distinguished hybrid fibers. Muscle fiber characteristics differed depending on species and muscle types.

• 한국식품과학회지
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• 제30권6호
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• pp.1409-1414
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• 1998

Zearalenone에 특이한 단크론성 항체를 개발하기위하여 형질세포종세포인 myeloma cell $(P3{\times}63Ag.V653)$과 zearalenone-oxime-bovine serum albumin (BSA) 결합체를 항원으로 면역시킨 BALB/c female mice의 비장세포를 융합시켜 hybridoma cell을 생산하였다. 그들의 항체가를 indirect competitive ELISA법으로 측정한 결과 zearalenone에 대하여 높은 친화력을 갖는 5세포주를 얻었다. 그중 Z-2-M26 hybridoma cell line이 생산하는 단크론성항체는 특히 zearalenone과 민감하게 반응하였고, ${\alpha}-zearalenol$과도 약간의 교차반응을 보였으나 ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ 및 DON과는 거의 반응하지 않았다. 따라서 본 실험에서 개발된 항체는 앞으로 zearalenone에 대한 민감하고 정량적인 효소면역측정법의 개발을 위한 중요한 면역시약으로 사용될 것으로 생각한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.150-154
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• 2002

Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. An indication of the emerging significance of antibody-based therapeutics is that over a third of the proteins currently undergoing clinical trials in the United States are antibodies. Until the late 1980's, antibody technology relied primarily on animal immunization and the expression of engineered antibodies. However, the development of methods for the expression of antibody fragments in bacteria and powerful techniques for screening combinatorial libraries, together with the accumulating structure-function data base of antibodies, have opened unlimited opportunities for the engineering of antibodies with tailor-made properties for specific applications. Antibodies of low immunogenicity, suitable for human therapy and in vivo diagnosis, can now be developed with relative ease. Here, antibody structure-function and antibody engineering technologies are described.