• Microbiology and Biotechnology Letters
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• v.36 no.1
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• pp.12-20
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• 2008

Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of MVM.

• Proceedings of the KSEEG Conference
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• 2003.04a
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• pp.76-79
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• 2003

국내의 토양오염 공정시험방법에서는 Zn, Ni 추출시 산분해법에 가까운 방법을 사용하는 반면, Cd, Cu, Pb, $Cr^{6+}$ 추출시 0.1N HCl용액으로 산처리하여 1시간을 진탕한 후 이를 필터로 여과하여 분석용액을 추출하는 용출법을 사용하고 있다(환경부, 2001). 시료내에는 완충 물질이 존재하기 때문에 용출법 사용시 초기 pH 인 1(0.1N HCl)이 유지되지 않아 완충능력이 높은 토양의 경우 현재 국내 공정법상의 용출법이 중금속 오염정도를 추정하는데 적절치 않을 수 있다. (중략)

• The Magazine of the Society of Air-Conditioning and Refrigerating Engineers of Korea
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• v.33 no.1
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• pp.11-21
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• 2004

실내공기질 공정시험법의 개요와 향후 연구진행 방향, 국내외의 실내공기오염물질에 대한 측정 규격의 종류 및 특성, 공정시험법 도출을 위한 기본적인 고려사항 등에 대하여 간단히 살펴보고자 한다.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.11a
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• pp.106-109
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• 2005

확산형 시료 채취기와 직독식 기기(공정시험법, Nitrogen Oxides Analyzer Model; EC 9841, Ecotech, Australia)에 의한 $NO_2$, 농도를 비교하고, 능동시료채취기(공정시험법)와 확산형포집기에 의한 HCHO(포름알데히드) 농도를 비교하기 위해 서울 ${\cdot}$ 경기 또는 대전, 충남 ${\cdot}$ 북지역에 소재한 11개 시설(종합병원 4곳, 노인 병원 1곳, 보건소 1곳, 복지관 3곳, 보육시설 2곳)을 대상으로 수행하였다. 1. 포름알데히드의능동 포집법(공정시험법)에 의한 시료(n=87)의 평균농도는 $11.44{\pm}11.07ppb$이고, 확산형 시료 채취기 의한 시료(n=40)의 평균농도는 $11.91{\pm}7.37ppb$으로 비슷한 값이 나왔고, 통계적으로 유의하지 않았다(p=0.806). 2. 포름알데히드 능동 포집법에 의한 농도와 확산형 시료 채취기에 의한 농도와의 상관계수 r=0.404(p=0.037)로 나타나 이 두 가지의 방법은 특정시간 포름알데히드 측정에 사용하여도 어느 정도 비교하기에는 적합할 것으로 생각된다. 3. 이산화질소의 노출정도는 직독식 기기(공정시험방법)와 확산형 시료 채취기로 각각 1시간 (오전, 오후 각각 2회), 8시간 측정하였다. 공정시험방법(n=61)에 의한 1시간-시료 평균농도는$44.48{\pm}37.96ppb$이고, 확산형 시료 채취기(n=61)에 의한 1시간-시료 평균농도는 $3.58{\pm}2.07ppb$으로 통계적으로 유의하였다(p=0.000). 직독식 기기(n=61)에 의한 8시간-시료 평균농도는 $34.85{\pm}22.83ppb$이고, 확산형 시료 채취기(n=61)에 의한 8시간-시료 평균농도 $8.32{\pm}4.44ppb$으로 통계적으로도 유의하였다(p=0.000). 4. 이산화질소를 직독식 기기(공정시험방법)와 확산형 시료 채취기로 측정한 1시간-시료 농도의 상관계수 r=0.253(p=0.268)이고 8시간-시료 일 때 상관계수 r=0.367(p=0.102)로 나타나 확산형 시료 채취기를 직독식 기기(공정시험방법) 대체 사용방법으로 이용하기에는 적합하지 않다고 생각된다.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.199-208
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• 2015

Biologics and medical devices manufactured with bovine-derived raw materials have the risk of viral contamination. Therefore, viral validation study is essential to ensure the safety of the products. Bovine adenovirus type-1 (BAdV-1) is one of the common bovine viral pathogens. For quantitative detection of BAdV-1 during the manufacture of biologics and medical devices, a TaqMan probe real-time PCR method was developed. Specific primers and TaqMan probe for amplifying and detecting BAdV-1 DNA were designed. Specificity, limit of detection (LOD), and robustness of the method was validated according to international guideline on the validation of nucleic acid amplification tests for the pathogen detection. The sensitivity of the assay was found to be $7.44{\times}10^1\;TCID_{50}/ml$. The real-time PCR method was reproducible, very specific to BAdV-1, and robust. Moreover, the method was successfully applied to the validation of Chinese Hamster Ovary (CHO)-K1 cells artificially infected with BAdV-1, a commercial CHO master bank, and bovine type 1 collagen. The overall results indicate that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BAdV-1 contamination during the manufacture of biologics and medical devices using bovine-derived raw materials.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.228-236
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• 2008

Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.

• Proceedings of the Korea Air Pollution Research Association Conference
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• 2002.04a
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• pp.141-142
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• 2002

대기환경 기준물질(criteria pollutants)은 NOx를 비롯하여 $O_3$, CO, SOx, PM 10, Pb 인데, 이러한 물질은 환경부에서 법적으로 규정한 대기오염공정 시험법에 따라서 분석된다. 대기오염공정 시험법이 충분한 과학적 근거를 갖는 분석법이 되기 위해서는 분석방법, 분석기기, 분석자 등에 의해 발생되는 불확실성을 최소화시키는 체계적인 관리가 필요하고, 분석기술의 발달수준, 측정분석의 요구수준, 새로운 국제 규격 통에 맞게 지속적으로 보완되어야 한다. (중략)

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.267-274
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• 2015

Biopharmaceuticals and the cell substrates used for their manufacture are currently tested for porcine adventitious viruses due to the widespread use of porcine trypsin in cell culture. Porcine transmissible gastroenteritis virus (PTGV) is one of the major adventitious porcine viruses causing contaminated during the manufacture of biopharmaceuticals. Therefore, rapid and sensitive detection of PTGV is essential in ensuring the safety of biopharmaceuticals. A TaqMan probe real-time RT-PCR method was developed for the quantitative detection of PTGV contamination in cell substrates, raw materials, manufacturing processes, and final products, as well as PTGV clearance validation. Specific primers for the amplification of PTGV RNA were selected, and PTGV RNA was quantified by use of a specific TaqMan probe. Specificity, limit of detection (LOD), and robustness of the method was validated according to international guidelines on the validation of nucleic acid amplification tests. The sensitivity of the assay was calculated to be 1.10 × 10⁰ TCID₅₀/ml. The real-time RT-PCR method was validated to be reproducible, very specific to PTGV, and robust. The established real-time RT-PCR assay was successfully applied to the validation of Chinese Hamster Ovary (CHO)-K1 cells artificially infected with PTGV.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.339-347
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• 2012

Viral safety is an important prerequisite for clinical preparations of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacturing process. In particular, Chinese hamster ovary (CHO) cells are highly susceptible to several RNA viruses including reovirus (Reo), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus (BPIV) and there have been reports of such viral contaminations. Therefore, viral detection during the CHO cell process is necessary to ensure the safety of biopharmaceuticals against viruses. In this study, a multiplex reverse transcription (RT)-PCR assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect Reo, BVDV, and BPIV during the manufacture of cell culture-derived biopharmaceuticals. Specific primers for Reo, BVDV, and BPIV were selected, and a multiplex RT-PCR was optimized. The sensitivity of the assay for simultaneous amplification of all viral target RNAs was $7.76{\times}10^2\;TCID_{50}/ml$ for Reo, $7.44{\times}10^1\;TCID_{50}/ml$ for BVDV, and $6.75{\times}10^1\;TCID_{50}/ml$ for BPIV. The multiplex RT-PCR was proven to be very specific to Reo, BVDV, and BPIV and was subsequently applied to the validation of CHO cells artificially infected with each virus. It could detect each viral RNA from CHO cells as well as culture supernatants. Therefore, it was concluded that the multiplex RT-PCR assay can be applied to detection of the adventitious viruses during the manufacture of cell culture-derived biopharmaceuticals.

• Journal of Food Hygiene and Safety
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• v.33 no.4
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• pp.296-305
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• 2018

Spiroxamine, one of fungicides, is used to control powdery mildew in various crops and black yellow sigatoka in bananas. The major strength of spiroxamine is to control powdery mildew in various crops and bananas yellow sigatoka in bananas. The compound has shown a high level of activity, good persistence and crop tolerance. Besides powdery mildew, good control of rust, net blotch and Rhynchosporium diseases been indicated in cereals, together with a complementary activity against Septoria diseases. In 2017, the maximum residue limit (MRL) of spiroxamine established in Korea. According to Ministry of ood and rug afety) regulations, spiroxamine residues defined only parent compound. Thus, a analytical method is needed to estimate the residue level of the parent compound. The objective of this study was to develop and validate analytical method for spiroxamine in representative agricultural commodities. Samples were extracted with acetonitrile and partitioned with dichloromethane to remove the interfering substances. The analyte were quantified and confirmed liquid chromatograph-tandem mass spectrometer (LC-MS/MS) in positive-ion mode using multiple reaction monitoring (MRM). Matrix matched calibration curves were linear over the calibration ranges ($0.0005{\sim}0.1{\mu}g/mL$) for the analyte in blank extract with coefficient of determination ($r^2$) > 0.99. For validation purposes, recovery studies will be carried out at three different concentration levels (LOQ, 10LOQ, and 50LOQ) performing five replicates at each level. The recoveries 70.6~104.6% with relative standard deviations (RSDs) less than 10%. All values were consistent with the criteria ranges in the Codex guidelines (CAC/GL40, 2003) and MFDS guidelines. proposed analytical method be used as an official analytical method in the Republic of Korea.