• Food Science and Biotechnology
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• 제17권3호
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• pp.547-552
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• 2008

The detection of carbamate (carbofuran, carbaryl, benfracarb, thiodicarb, and methomil) and organophosphate (diazinon, cadusafos, ethoprofos, parathion-methyl, and chlorpyrifos) pesticide residues with very low detection limits was carried out using surface plasmon resonance (SPR) based equipment. The capacity to develop a portable SPR biosensor for food safety was also investigated. The applied ligand for the immunoassays was polyclonal goat anti-rabbit immunoglobulin (IgG) peroxidase conjugate. Concentration tests using direct binding assays showed the possibility of quantitative analysis. For ligand fishing to find a proper antibody to respond to each pesticide, acetylcholinesterase (AChE), and glutathione-S-transferase (GST) were tested. The reproducibility and precision of SPR measurements were evaluated. With this approach, the limit of detection for pesticide residues was 1 ng/mL and analysis took less than 11 min. Thus, it was demonstrated that detecting multi-class pesticide residues using SPR and IgG antibodies provides enough sensitivity and speed for use in portable SPR biosensors.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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• pp.690-695
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• 2014

Bacterial 1-aminocyclopropane-1-carboxlyate (ACC) deaminase (AcdS) is an enzyme that cleaves ACC, a precursor of the plant hormone ethylene, into ${\alpha}$-ketobutyrate and ammonia. The acdS gene was cloned from Pseudomonas fluorescens, which was capable of improving the seedling of Chinese cabbage under salinity condition. The recombinant AcdS (rAcdS) exhibited optimal activity at pH 8.5 and $30^{\circ}C$. Strong activity was sustained at up to 100 mM NaCl. The polyclonal anti-P. fluorescens AcdS antibody was produced in a rabbit that had been immunized with the purified rAcdS. This antibody successfully recognized the homologous antigens derived from the total proteins of isolated plant growth-promoting microorganisms. A statistically significant correlation was observed between the intensity of hybridization signal and AcdS activity measured by a biochemical method, suggesting its application as a useful indicator for active deaminases.

• Journal of Microbiology
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• 제35권2호
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• pp.87-91
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• 1997

Monoclonal antibodies were prepared in order to an assay method for Vibrio vulnificus. Sixteen mouse ybridoma cell lines were established by immunization of whole cell antigen to BALB/c mice, fusion with SP2/O myeloma cells, and cloning. Most of them secreted IgM.lambda. antibodies. A sandwich enzyme-linked immunosorbent assay was developed with rabbit anti-V. vulnificus polyclonal antibodies as capture antibody, an IgM monoclonal antibody as detector antibody, and goat anti-mouse IgM-alkaline phosphatase conjugate as developer antibody. The range of detection was 10$\^$4/ to 10 V. vulnificus cells per microplate well. When four related Vibrio species were tested for cross-reactions, V. parahaemolyticus showed 3.5% reactively and V. carchariae, V. fluvialis, and V. furnisii showed negligibal (<1%) cross-reactivity.

• 생명과학회지
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• 제19권2호
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• pp.284-288
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• 2009

선천성 면역은 숙주의 물리적 방어벽을 뚫고 침입하는 감염성 질병 원인균에 대항하는 첫 번째 방어로서 아주 중요한 역할을 한다. Mannose-binding lectin (MBL 또는 mannan-binding protein, MBP)은 혈청 내에 존재하는 면역성 단백질로서 감염 후 즉시 유발되는 acute phase response의 특정 단백질이다. MBL 단백질은 세균, 바이러스, 곰팡이, 기생충 등의 탄수화합물 구조에 결합하여 식균 작용을 돕거나 보체경로를 활성화 시킨다. MBL 단백질은 C-말단이 탄수화물을 인식하는 도메인이며, 연결 목 부위와 콜라겐 부위로 구성되어 있다. 혈청 내의 MBL 농도가 낮으면 높은 빈도로 면역결핍현상이 관찰된다고 알려져 있다. MBL 단백질의 기능과 유전에 대해 많은 연구가 되어져 왔으나 아직 MBL 단백질 복합체 등에 대한 연구는 많이 이루어져 있지 않다. 따라서 MBL 연구에 필수적인 MBL cDNA 제조와 재조합 단백질의 합성, 그리고 재조합 단백질을 항원으로 사용하여 polyclonal antibody를 생산한 연구 결과를 보고하는 바이다. 본 연구결과로 획득한 MBL cDNA, 재조합 단백질과 anti-MBL 항체는 앞으로의 MBL 연구에 절대적으로 필요한 도구가 될 것으로 생각된다.

• 대한수의학회지
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• 제30권2호
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• pp.215-221
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• 1990

The present study was intended to use the peroxidase-antiperoxidase method for the identification of hog cholera virus(HCV) in the lymphatic organs of HCV-infected pigs. Sections were incubated with primary antibody (rabbit anti-HCV polyclonal or mouse anti-HCV monoclonal), followed by incubation with linkserum (goat anti-rabbit IgG) in excess and rabbit or mouse PAP complex. The viral antigen was localized mainly in the cytoplasms of lymphoid cells and macrophages. Positive reaction cells were frequently detected in the marginal areas of the germinal centers of the spleens, and also found in the tensils and lymph nodes. The method approved to be highly specific for the identification of the virus and allowed a precise localization of the viral antigen in infected cells.

• Journal of Applied Biological Chemistry
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• 제48권1호
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• pp.16-25
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• 2005

To develop an enzyme-linked immunosorbent assay (ELISA) for the detection of the residues of the acaricide fenazaquin, five haptens were synthesized and assessed. A competitive indirect format was used with polyclonal antibodies. Under an optimized condition using the selected rabbit C antiserum, an $IC_{50}$ of $96.97\;ng{\cdot}ml^{-1}$, the detection range of $14.9{\sim}631\;ng{\cdot}ml^{-1}$, and the lowest detection limit of $8\;ng{\cdot}ml^{-1}$ were obtained. Some structurally related compounds of practical use showed low crossreactivities to the antibody. Highest cross-reactivity observed with hapten IV indicates that the antiserum C recognizes very well quinazoline ring, 4-tert-butylphenyl, and an adequate length of spacer arm. The length of spacer arm affected recognition of quinazoline ring and 4-tert-butylphenyl moieties. When applied to apple and pear, recoveries were within acceptable ranges of $93.18{\sim}104.77%$ (n = 4) and $79.40{\sim}111.95%$ (n = 4), respectively.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권1호
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• pp.31-41
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• 2002

Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

• 생약학회지
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• 제46권4호
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• pp.279-282
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• 2015

In order to quantify compound K(CK), anticancer component of Panax ginseng C. A. Meyer, high titer rabbit polyclonal antibodies (pAbs) were raised against a conjugate of CK and bovine serum albumin coupled by a periodate oxidation method. Coating antigen (CK-OVA) was also prepared by the same method with OVA. As a result of optimization of antiserum dilution (2,000 fold), coating antigen ($25{\mu}g/ml$) and other condition (incubation time, temperature and washing method), ELISA method for the determination of CK was established. The measuring range extended from 0.5 ng/ml to 25 ng/ml of CK. The antibodies exhibited minor or even no cross reactivities with protopanaxatriol (1.56%) and other tested ginsenosides, $GRb_1$ (0.11%), $GRg_1$ (0.07%) except protopanaxadiol (87.2%) from the structural similarity. And the antibody showed good correlation (r=0.987) between the assay values obtained by this ELISA method and HPLC. Therefore, the ELISA method could be very useful tools for the determination of CK in biological fluids because of their high sensitivity and specificity.

• The Journal of Korean Physical Therapy
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• 제19권4호
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• pp.33-41
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• 2007

Purpose: To investigate environmental enrichment and nerve stimulation follows in application times with the change of BDNF & Trk-B receptor in the motor cortex and spinal cord. Methods: Experimental groups were divided into the five groups. Group I: normal control group, Group II: experiment control group, Group III: sciatic never electrical stimulation after MCAO, Group IV: application of only environmental enrichment after MCAO, Group V: never electrical stimulation with environmental enrichment after MCAO. Histologic observation and coronal sections were processed individually in goat polyclonal antibody phosphorylated BDNF and rabbit polyclonal antibody Trk-B receptor. Results: In immunohistochemistric response of BDNF and Trk-B, group II were showed that lower response effect at postischemic 1 days, 3 days, and 7 days. Group V were showed that increase response effect at postischemic 3 days, 7 days and 14 days. Specially showed that the most response effect at postischemic 14 days. In neurobehavioral assessment, group V were significantly difference from other groups on between-subject effects. Conclusion: The above results suggest that combined environmental enrichment with peripheral nerve electrical stimulation in focal ischemic brain injury were more improved that the change of BDNF & Trk-B receptor expression than non treatment.

• Parasites, Hosts and Diseases
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• 제33권3호
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• pp.155-164
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• 1995

Cyptosporidium parvum의 발달 단계별 actin과 myosin의 분포 위치를 면역황금염색법을 이용하여 관찰하였다. $Depomedrol^{\circledR}$을 ICR마우스에 피하주사하여 면역억제시킨 후 C. parvum이 발현된 마우스 회장을 잘라 LR gold로 포매하여 초박절편을 떴다. 일차항체로는 chickenbackmuscle actin과 bovine uterus myosin에 대한 rabbit polyclonal antibody를 사용하였고 이차항체로는 10 mm 크기의 황금입자가 결합된 goatanti-rabbit lgG를 반응시켰다 Uranylacetate와 leadcitrate로 염색한 후 투과전자현미경으로 관찰하였다 Trophozoite에서는 세포막에서 주로 actin과 myosin이 관찰되었고 feederorganelle 주위 세포질에는 actin이 분포하였다. Meront와 같이 활발히 분열하고 있는 단계에서는 세포막과 세포질 전체에 actin이 분포되어있었으며 myosin은 세포막에서만 소량 관찰되었다. 핵과 anlage of rhoptries 등은 두 단백질에 모두 염색되지 않았다. Macrogametocyte 에서는 amylopectin-lile bodies에서 actin과 myosin이 모두 관찰되었으나 wall forming bodies에서는 관찰되지 않았고 feederorganelle 주위 세포질 부분에서는 actin이 관찰되었다. Sporozoite를 포함하는 oocyst와 merozoite를 포함하는 meront에서는 세포막과 세포막사이에서 actin이 다수 관찰 되었으며 myosin은 소량 관찰되었다. Merozoites가 빠져나가 속이 비어있는 parasitophorous vacuole중에는 microspike를 형성한 것들이 종종 관찰되었고 이것이 좀더 길어져 마치 microvilli와 같이 보이는 경우도 있었으며 이러한 구조물에서도 actin이 다수 관찰되었다. 이상의 결과로 미루어 actin과 myosin은 세포막에 주로 분포하면서 C. parvum의 형태를 유지시키며 또한 세포막의 움직임을 조절하는 cytoskeletalproteiA으로서의 역할이 주된 작용일 것으로 생각되었다.