효소면역측정법을 이용한 Fumonisin의 검출법 개발

손동화,한성민,임선희,이인원,조선희,강신영,이경애 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

Fumonisin에 대한 효소면역측정법(ELISA)을 개발하기 위하여 특이항체를 생산하고, 분석조건을 확립하여, 인위적으로 오염시킨 옥수수 시료의 분석을 행하였다. Fumonisin B_1 (FB_1)을 cholera toxin (CT)에 결합시킨 FB_1-CT conjugate를 면역원으로 하여 Freund's adjuvant와 함께, 또는 단독으로 두 군의 토끼에 면역하고 특이항체를 생산하였다. 항혈청을 ELISA로 분석한 결과 그 adjuvant와 함께 면역한 경우 항체역가(titer)가 높게 나타났다. 가장 높은 항체역가(1 : 16,000)를 나타낸 항혈청 및 그로부터 정제한 항체를 이용하여 간접법 및 직접법에 의한 경합적 ELISA(ciELISA 및 cdELISA)를 각각 확립하였다. 이 항체의 유사독소와의 교차반응을 ciELISA로 조사하였을 때, FB_3에 대한 교차반응율은 2%로 매우 낮았으나, FB_2에 대한 것은 179%로 다소 높았다. FB_1의 검출한계는 ciELISA에서 0.03ppb, cdELISA에서 0.3 ppb로 각각 나타났다. 실제 곡물시료의 분석에 ELISA 활용가능성을 조사하기 위하여 인위적으로 FB_1을 오염시킨 옥수수를 75% methanol로 추출하고 그 회수율을 측정한 결과, 분석이 불안정하여 회수율이 일정하게 나타나지 않았다. 그러나 방해물질의 제거를 위하여 시료추출액을 strong anion exchange(SAX) cartridge를 거쳐 세척한 다음 분석하였을 때, 3~10 ppm 범위의 FB_1 오염 옥수수에서 평균 34%(CV의 평균, 8.2%)의 안정된 회수율을 보였다. In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishement of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin B_1 (FB_1) conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1 : 16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigate by the ciELISA is was very low against B_3 (2%) but high against fumonisin B_2 (179%). The sensitivity of the ELISAs was also very high, because the detection limit for FB_1 was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of FB_1 was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of FB_1 from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

효소면역측정법에 의한 우유중의 Aflatoxin M1 분석

손동화,임선희,이인원 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

국내산 우유중 aflatoxin M_1 (AFM_1)의 오염현황을 조사하기 위하여, 효소면역측정법(ELISA)을 개발하고 이를 이용한 독소의 정량을 행하였다. 항체의 생산을 위하여 AFM_1 bovine serum albumin 결합체를 Freund's adjuvant와 함께 토끼에 피하주사하여 면역하였다. 가장 높은 항체역가를 보인 항혈청과 AFB_1-horseradish peroxidase 결합체를 이용하여 직접법에 의한 경합적 효소면역측정법(competitive direct ELISA ; cdELISA)의 조건을 확립하였으며, 그 검출한계는 0.003ppb로 나타났다. 또한 유사독소에 대한 교차반응율은 aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a 에 대하여 각각 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, 및 0%이었다. 다음으로, AFM_1을 인위적으로 오염시킨 우유를 C_18 cartridge로 세척후 cdELISA로 분석하였을 때, 0.01~1 ppb(10~1,000 ppt)의 범위에서 그 회수율의 평균은 104%(CV의 평균, 6.4%)로 나타났다. 시유 및 목장우유를 대상으로 AFM_1의 오염현황을 cdELISA로 조사하였을 때, 평균 오염도는 80.4±55.0 ppt (n=64; range, 5,6~280 ppt)이었다. For a survey of the occurrence of aflatoxin M_1 (AFM_1) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM_1 conjugated to bovine serum albumin (AFM_1-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB_1-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM_1 whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM_1 and followed by cleanup with C_18 cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01~1 ppb (10~1,000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM_1 was 80.4±55.0 ppt (n=64; range, 5,6~280 ppt).

느타리버섯 세균성갈빈병균 Pseudomonas tolaasii의 효소면역검출법

이향범,전낙범,손동화,유승헌 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

느타리버섯의 세균성갈반병균인 P. talaasii(PT)를 신속 간편하게 검출할 수 있는 효소면역측정법(ELISA)을 개발하였다. 특이항체를 생산하기 위하여 PT(5×10 exp (7) cfu)를 Freund's adjuvant와 함께 토끼에 수차례 면역하였으며, 가장 높은 역가를 나타낸 항혈청을 이용하여 비경합 ELISA 및 경합 ELISA를 각각 확립하였다. 표준곡선으로부터 PT의 그 검출한계는 각각 2×10 exp (2) cfu/ml 및 3×10 exp (2) cfu/ml가량으로 나타났다. 교차반응은 비경합 ELISA의 경우 P. agarici, P. reactans 및 비병원성 Pseudomonas속 균주와 1/10^3이하의 매우 미약한 반응을 보였으나 P. salanacearum, Erwinia chrysanthemi, Streptococcus mutans, Xanthomonas citri 및 곰팡이인 Fusarium oxysporum 등 다른 균주와의 반응성이 거의 없었으며, 경합 ELISA의 경우 PT이외에는 같은 속 균주에 대하여도 반응성이 거의 없었다. ELISA를 이용하여 버섯으로부터 분리된 18균주에 대하여 PT의 여부를 조사하였을 때 병원성과 white line 반응성을 동시에 나타내었던 11균주에 대하여만 명확한 양성으로 나타나, 본 연구에서 확립한 ELISA는 간편하고 정확한 PT검출법임이 확인되었다. For simple and rapid detection of Pseudomonas tolaasii (PT), a bacterial brown blotch pathogen of oyster mushroom, enzyme-linked immunosorbent assays (ELISA) were developed. To produce specific antibody, PT (5 × 10 exp (7) cfu) and Freund's adjuvant were subcutaneously immunized into rabbits several times. By using the antiserum showing the highest titer, we established noncompetitive and competitive ELISA's. Standard curves of the ELISA's showed that the detection limits were 2 × 10 exp (2) cfu/ml and 3 × 10 exp (2) cfu/ml, respectively. When investigated by noncompetitive ELISA, cross reactivities of the anti-PT antibodies against P. agarici, P. reactans, and other fluorescent Pseudomonas spp. were very low (<1/10^3), but those against P. solanacearum, Erwinia chrysanthemi, Streptococcus mutans, Xanthomonas citri, and a fungus Fusarium oxysporum were almost none. However, when investigated by competitive ELISA, the reactivities against any other strains except PT were almost none. When the ELISA's were applied to 18 strains derived from mushrooms in order to identify PT, only 11 strains showing both pathogenicity and white line reactivity were obviously positive. These results showed that the ELISA's could be convenient tools to detect PT in accordance with existing methods.

미용업 종사자들의 피부, 호흡기 및 근골격계 자각증상에 관한 유병률

이종태,박봉진,엄상화,김성준,강동묵,손혜숙,정귀원,강민숙,박성희 大韓産業醫學會 1999 대한직업환경의학회지 Vol.11 No.3

Objectives: Present study was conducted to evaluate work-related symptom prevalence among hairdressers. Methods: Exposed group comprised 184 employee employed 73 hair salons in 6 district of Pusan city, and non-exposed group comprised 119 people living recent apartments. A trained interviewer interviewed them with organised questionnaire which included dermatologic, respiratory and musculo-skeletal symptoms. Results : Prevalence of hand eczema was 28.3% in exposed group, and 5.8% in non-exposed group. Adjusted odds ratio for age and atopy history was 4.30(2.34-7.93). Prevalence of respiratory symptom (coughing) in exposed group was 22.1%, and 9.4% for non-exposed group. Adjusted odds ratio for coughing which was adjusted for age, smoking and atopy history was 2.76(1.32∼5.78). Prevalences of musculo-skeletal symptoms among exposed group were neck(59.9%), shoulder(76.6%), upper back(41.2%), lower back(72.2%), arm and elbow(31.3%), wrist(44.2%), finger(35.0%), leg(71.1%). Adjusted Odds Ratios for musculo-skeletal symptoms which was adjusted for age were neck 2.13(1.29∼3.51), shoulder 2.52(1.50∼4.24), upper back 1.71(1.01∼2.88), lower back1.78(1.06∼2.99), arm and elbow 3.10(1.62∼5.94), wrist 2.09(1.23∼3.57), finger 4.83(2.41∼9.68), leg 3.46(2.07∼5.79). Conclusions : These results show that employees in hair salon are likely to have high risk for work-related dermatologic, respiratory and musculo-skeletal symptoms and diseases. Hence, prevention methods from those work-related diseases are required to be developed. Also, the scope of occupational and environmental medicine should be expanded to service area including hairdressers.

Development of Anti-Allergic Functional Food Using Natural Materials

Dong-Hwa Shon,Hee Soon Shin 한국식품영양과학회 2018 한국식품영양과학회 학술대회발표집 Vol.2018 No.10

Enzyme-linked Immunosorbent Assay for the Detection of Hen's Egg Proteins in Processed Foods

Shon, Dong-Hwa,Kim, Hyun-Jung,Kim, Soo-Ho,Kwak, Bo-Yeon Korean Society for Food Science of Animal Resource 2010 한국축산식품학회지 Vol.30 No.1

The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

Strategy and Outline of Anti-Allergic Food Development

Dong-Hwa Shon 한국식품영양과학회 2016 한국식품영양과학회 학술대회발표집 Vol.2016 No.10

Study on the Transfer Functions for Detecting Windings Displacement of Power Transformers with Impulse Method

Shon, Chae-Hwa,Yi, Sang-Hwa,Lee, Heun-Jin,Kang, Dong-Sik The Korean Institute of Electrical Engineers 2012 Journal of Electrical Engineering & Technology Vol.7 No.6

The paper investigates three types of transfer function methods for detecting displacements of winding in a model transformer. To acquire these transfer functions, the measuring method of input voltage, current and its response is used in impulse method. The applied impulse voltages had three rising times, which were short rising time (less than 0.6 ${\mu}s$), medium rising time (about 0.8 ${\mu}s$) and long rising time (about 1 ${\mu}s$) in front waves. Every 10 measurements of voltage and current waves were averaged from 50 measurements of voltage and current waves. These transfer functions were tested in normal, 24mm elevated and 48mm elevated windings conditions and were analyzed with correlation coefficients and spectrum deviations. In the analysis, the results depend on the types of transfer functions and the rising times of input voltages.

ELISA System for aflatoxin M1 in cow's milk without cleanup procedure

Dong Hwa Shon,Sun Hee Lim,Hyang Burm Lee and Yin Won Lee 한국식품과학회 1996 한국식품과학회 학술대회 Vol.1996 No.1

Enzyme-Linked Immunosorbent Assay for the Detection of Hen`s Egg Proteins in processed Foods

Dong Hwa Shon,Hyun Jung Kim,Soo Ho Kim,Bo Yeon Kwak 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.1

The Hen`s egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries (mean±SD) were 129±13.7%, 73.9±12.5%, and 65.5±13.6%, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen`s egg proteins in processed foods.