• BMB Reports
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• 제32권5호
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• pp.474-479
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• 1999

Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

• 대한소아치과학회지
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• 제36권4호
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• pp.522-530
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• 2009

Monoclonal antibody를 이용한 Saliva-$check^{TM}$ Mutans키트의 타액 Streptococcus mutans검출방법으로서의 활용도와 임상적 우식지수 및 기존의 세균배양방법과의 상관관계를 알아보기 위해 2008년 2월에서 5월 중 평촌키즈웰치과에 내원한 만 2세에서 만 8세 사이의 92명의 아동을 대상으로 Streptococcus mutans 검출검사를 실시하였으며 또한 우식에 영향을 미치는 다른 요인인 치면세균막 pH와 타액완충능력검사를 실시하여 다음과 같은 결론을 얻었다. 1. Saliva-$check^{TM}$ Mutans 검사결과 양성을 나타낸 아동은 27명으로 29.3%이었고, 음성을 나타낸 아동은 65명으로 70.65%이었다. 우식경험유치면률은 음성 아동 13.89%, 양성 아동 25.23%으로 나타났다. 2. Monoclonal antibody를 이용한 검사방법 인 Saliva-$check^{TM}$ Mutans와 기존의 세균 배양방법인 $Dentocult^{(R)}$-SM은 각각 상이한 검사방법을 이용함에도 불구하고 검사결과 높은 상관관계를 보였다(p<0.01). 3. Saliva-$check^{TM}$ Mutans검사와 치면세균막 pH검사와는 역상관관계를 나타내었으나(p<0.01),타액완충능 검사와는 상관관계가 나타나지 않았다(p>0.05). 이상의 결과로 보았을 때 Monoclonal antibody를 이용한 검사방법인 Saliva-$check^{TM}$ Mutans는 구강 내 Streptococcus mutans를 측정하는 방법으로 적당하며 또한 검사에 필요한 시간을 대폭 줄일 수 있고 사용방법도 매우 간편하게 개발되어 환자들에게 적용함에 있어 효과적이라고 사료되었다.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1152-1161
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• 2007

An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.

• 한국가축번식학회지
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• 제14권3호
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• pp.165-173
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• 1990

A simple and sensitive microplate enzyme immunoassay(ELISA) was developed for progesterone, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxise(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5pg-120pg/well and intra- and inter assay coefficients of variation for progesterone standard curve(0.1ng-3.2ng/ml) were ranged 4.4-10.6% and 5.-12.6%, respectively. They assay is performed in less than two hours and provide reliable values to differentiate among samples from day 0(A.I.), day 14 and day 19. The discriminatory levels for early pregnancy diagnosis are [>10ng(day 19) & decreasing rate <1.5 : pregnancy] and [ 7ng & decreasing rate 1.5 : non-pregnancy]. The accuracy of the pregnancy diagnosis for cows classified as positive(pregnancy) and negative(non-pregnancy) were 96% and 100%, respectively.

• 한국동물위생학회지
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• 제33권1호
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• pp.1-6
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• 2010

Monoclonal antibody (mAb)-based enzyme immune-slide assay (EISA) was used for the detection of Peste des petits ruminants (PPR) virus from field samples collected from a natural outbreak. The clinicopathological study was undertaken to diagnose the case primarily of PPR. Antigen was detected from discharges and faeces of infected goats and swabs of postmortem lesions prepared on glass slide or glass plate using acetone fixation. Nasal discharge collected at the early stage of disease course or lung is an appropriate ante- or postmortem sample for this technique, respectively. Convalescent polyclonal sera collected from recovered animals which were diagnosed as PPR by EISA showed high antibody titer against PPR by C-ELISA, demonstrating the satisfactory specificity of the test. Therefore, EISA is a sensitive and specific assay to confirm PPR infection both in field and laboratory conditions and especially suitable for developing country.

• 대한소아치과학회지
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• 제37권2호
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• pp.186-192
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• 2010

Streptococcus mutans는 구강 내에 상존하는 치아우식증의 주요 원인균으로서 치면의 피막에 부착 후 glucan을 형성하여 세균의 군락을 이루며, 외부로부터 공급된 자당대사를 통하여 유기산을 생성함으로써 법랑질을 탈회시킨다. 치아우식 활성도의 평가를 위한 단클론항체를 이용한 방법은 진료실에서 빠른 시간 내에 간편하게 타액에 존재하는 Streptococcus mutans의 정량분석이 가능한 방법이다. 이 연구는 3세에서 6세 사이의 어린이 15명을 대상으로 자극성 타액을 채취하여 시판 중인 단클론항체를 이용한 Salivacheck$^{TM}$ Mutans, strip을 이용한 Dentocult$^{(R)}$-SM 그리고 MSB배지 배양법으로서 타액 내 Streptococcus mutans를 측정한 후 그 값을 우식경험치아수와 비교하여 상관관계를 알아보았다. Saliva-check$^{TM}$ Mutans를 이용한 방법은 Dentocult$^{(R)}$-SM과 MSB배지법과 통계학적으로 유의한 상관관계를 보였으나(p<0.05), MSB배지법은 어린이의 우식경험치아수와 통계학적으로 유의한 결과를 나타내지 않았다(p=0.34).

• Clinical and Experimental Reproductive Medicine
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• 제21권1호
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• pp.89-97
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• 1994

These studies were carried out to investigate the properties of H-Y antigen purified by immunoaffinity chromatography using monoclonal H-Y antibody. Immunoaffinity column was prepared by the coupling of monoclonal antibody to the Aminolink Coupling Gel. Murine testis supernatant was applied onto the column and eluted by O.lM glycine-HCl buffer and 31${\mu}g$ of H-Y Ag was eluted from one testis. Purified H-Y Ag strongly reacted with Con A and lentil from 6 different kinds of lectins tested, which may indicate that sugar moiety of H-Y Ag is composed of glucose, mannose and their derivatives. Con A-sepharose affinity column was used to purified H-Y Ag based on that H-Y Ag is glycoprotein. The fraction eluted by 0.2M Me-${\alpha}$-D-mannoside from the column loaded with murine testis supernatant was identified to be H-Y Ag by dot blot test. Molecular weight of the purified H-Y Ag was estimated by Sepharose G-75 gel filtration and SDS-PAGE, and showing that it was about 67,000 dalton. In fluorescence test, the ratio of XY embryos and XX embryos was 1:1.

• 한국발생생물학회지:발생과생식
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• 제9권2호
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• pp.135-142
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• 2005

무척추동물에서 척추동물에 이르기까지 대부분의 난생동물들의 난황 단백질의 전구체를 vitellogenin(VTG)이라 한다. 난생 척추동물에서 VTG는 간에서 합성되어 혈액을 통해 난세포로 전이된다. 암컷 어류는 정상적인 생식주기에서 난황단백전구체 형성이 시작되면 혈중 농도는 급격히 증가하게 된다. 수컷도 VTG의 유전자를 가지고 있기 때문에 낮은 수준의 내인성 에스트로겐으로 아주 적은 양의 단백질이 있을 수 있다. 그렇지만 수컷에 외인성에스트로겐 유사물질에 노출되면 그 양은 증가하게 된다. 따라서 어류에서 VTG는 내분비계 장애물질 효과를 조사하는 유용한 생물학적 추적자로 사용할 수 있다. 이 연구에서는 에스트로겐 유사물질에 오염된 지역에 서식하는 망둑어류의 혈중 VTG의 수준을 정량할 수 있는 방법을 제공하고자 한다. 이러한 목적을 위해 꾹저구(Chaenogobius annularis)의 VTG를 분리하고, 이에 대한 단클론 항체와 다클론 항체를 제작하여 혈중 VTG 수준을 정량할 수 있는 sandwich 경합 ELISA system을 만들었다. 난황 단백질에 대한 단클론 항체와 다클론 항체를 사용한 ELISA system에 대한 타당성 검토를 하였다. 순차적으로 희석한 암 컷의 혈청에 대한 흡광도 곡선은 VTG 표준농도의 곡선과 평행하였으나 수컷의 혈청에 대한 흡광도 곡선은 평행하지 않았다. 이 VTG에 대한 sandwich ELISA system으로 꾹저구(C. annularis)의 혈중 VTG의 수준을 정량뿐만 아니라 문절망둑(Acanthogobius flaviman)과 풀망둑(A. hasta)의 혈중 VTG 수준을 정량할 수 있다.

• 한국미생물·생명공학회지
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• 제46권3호
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• pp.269-276
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• 2018

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

• BMB Reports
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• 제29권3호
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.