• 제목/요약/키워드: A-fetoprotein

검색결과 99건 처리시간 0.02초

인간 $\alpha$-fetoprotein (AFP)에 대한 폴리클로날 항체의 생산 및 $\alpha$-fetoprotein 측정용 효소면역분석법 (competitive ELISA)의 개발 (Production of Polyclonal Antibody against $\alpha$-Fetoprotein and Polyclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for $\alpha$-Fetoprotein)

  • Michung Yoon
    • 대한의생명과학회지
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    • 제3권2호
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    • pp.115-123
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    • 1997
  • 인간 $\alpha$-fetoprotein (AFP)은 간암, 위암, 생식기 종양 및 신경관 이상인 환자를 검사하고 진단하는데 유용한 지표로 알려져 있다. 본 연구에서는 사람의 AFP를 분리정제 하여 폴리클로날 항체를 생산하고 인간 혈장과 양수내의 AFP를 측정하기 위한 경쟁적 효소면역 분석법을 개발하고자 하였다. 친화크로마토그래피법과 SDS-polyacrylamide 전기영동법을 이 용하여 양수로부터 AFP를 분리하였다. 정제된 AFP를 토끼에 주사하여 폴리클로날 항체를 생산하였으며, 이중면역확산법과 Western blot 분석법을 사용하여 본 연구실에서 제조된 항체의 항원 특이성이 대단히 높음을 확인하였다. AFP와 항혈청을 이용하여 표준곡선을 얻었으며, 민감도는 5ng/ml이었고, 작용범위는 5~l,000ng/ml이었다. 분석내 CV는 4.5%이었고, 분석간 CV는 8.5%이었다. 따라서 이러한 결과로 보아 본 연구에서 개발된 경쟁적 효소면역분석법이 AFP를 측정하기에 적절하며, 간암 등의 기초연구에도 많은 기여를 할 것으로 생각된다.

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吸着 크로마토그라피法에 의한 사람 ALpha-Fetoprotein의 分離 (Affinity Purification of Human Alpha-Fetoprotein)

  • Kang, Shin-Sung;Bang, Ok-Sun;Park, Tae-Kyu
    • 한국동물학회지
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    • 제29권4호
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    • pp.283-293
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    • 1986
  • 사람의 alpha-fetoprotein(AFP)에 대한 모노클론 항체의 생산 및 분석을 위하여, 태아조직을 재료로 추출법, DEAE-cellulose 및 concanavalin A-Sepharose, Cibacron blue F3GA-agarose, immunoadsorbent column등의 흡착크로마토그라피법에 의해 AFP를 분리하였다. 총 534g의 태아조직에서 AFP의 量은 8.76 mg으로서 순수분리되었음을 확인하였다.

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건강 검진자들의 혈청 Alpha-Fetoprotein 농도 (Serum Alpha-Fetoprotein Levels in Health Checkup Adults)

  • 김유현
    • 대한임상검사과학회지
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    • 제40권2호
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    • pp.86-93
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    • 2008
  • This study was performed to investigation of the serum alpha-fetoprotein (AFP) levels in healthy adults. A total of 2,160 (male 1,415, female 745) health checkup adults were examined for AFP levels by chemiluminescence immunoassay (CLIA) method, during the period from September, 2007 to August, 2008. The mean serum AFP level was 2.168 (0.605~20.35) ng/mL, and it was 2.309 (0.605~20.35) ng/mL in male, 1.906 (0.605~10.36) ng/mL in female, respectively. 1,816 (male 1,109, female 709) healthy adults were screened for the absence of viral hepatitis and normal alanine amino transferase (ALT) levels. The mean serum AFP level of healthy adult was 2.041 (0.605~7.83) ng/mL, and it was 2.181 (0.605~7.83) ng/mL in male, 1.822 (0.605~6.55) ng/mL in female, respectively. Serum AFP increased with age group, there was a higher level in male compared to female. These results suggests that the use of reference value of AFP in healthy adults in the Jeonbuk. With the reference value now defined, it becomes possible to compare levels in different populations.

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Alpha-fetoprotein 측정용 국산 방사면역측정법 시약의 평가 (The Evaluation of Domestic Immunoradiometric Assay Kit for Alpha-fetoprotein)

  • 원경숙;류진숙;문대혁;이희경
    • 대한핵의학회지
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    • 제34권4호
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    • pp.353-359
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    • 2000
  • 목적: 최근 국내에서 방사면역측정법용 ${\alpha}$-FP 검사시약(Riakey AFP IRMA $CT^R$, Sin-Jin Medics, Seoul, Korea)이 개발되어 그 성능을 평가해 보고자 본 연구를 시행하였다. 대상 및 방법: 검사방법은 시약 내 설명서의 권고사항을 준수하여 실시하였으며, 정밀도, 회수율, 평행성, 민감도에 대한 검사를 시행하였고, 다른 상업용 방사면역측정법용 a-FP 검사시약에 의한 측정값과의 상관관계를 조사하였다. 결과: 측정 내 정밀도는 세 준위의 혈청을 각각 10개의 튜브에 분주한 후 한 배치에서 검사하였는데, 평균 18.9 ng/ml에서 변이계수 5.3%, 평균 133 ng/ml에서 3.4%, 평균 332 ng/ml에서 변이계수 1.6%였다. 측정간 정밀도는 세 준위의 혈청을 하루 간격으로 5회 측정하여, 평균 20.9 ng/ml에서 변이계수 9.7%, 평균 137 ng/ml에서 3.2%, 평균 330 ng/ml에서 변이계수 4.1%였다. 회수율은 저, 중, 고 세 가지 농도의 혈청에 각각 시약 내 표준용액 10, 70, 250 ng/ml을 1:1로 섞은 후 검사하여, 17.6 ng/ml에서 각각 102%, 105%, 101%, 105 ng/ml에서 각각 103%, 95%, 105%였고, 245 ng/ml에서 각각 97.3%, 92.1%, 98.6%이었다. 평행성 검사는 세 개의 고농도 검체를 각각 2, 4, 8, 10, 20, 100 배 희석하여 한번에 검사하였으며, 평행성 검사에서 계산한 수식은 각각 y=0.98x-10.3, y=0.99x-8.32, y=0.98x-11.3이었다. 민감도는 측정가능한 최소농도가 0.63 ng/ml이었다. 175명의 환자검체의 측정결과를 다른 시약들로 측정한 것과 비교하였을 때 강한 상관관계를 보였다(r=0.98). 결론: 이상의 결과로 ${\alpha}$-FP 검사시약인 Riakey AFP IRMA $CT^R$은 임상에 적용할 수 있는 유용한 시약으로 판단된다.

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Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • 대한의생명과학회지
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    • 제7권3호
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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TCP10L negatively regulates alpha-fetoprotein expression in hepatocellular carcinoma

  • Shen, Suqin;Feng, Huan;Liu, Longjiang;Su, Wei;Yu, Long;Wu, Jiaxue
    • BMB Reports
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    • 제53권8호
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    • pp.431-436
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    • 2020
  • Alpha-fetoprotein (AFP) is one of the most commonly used and reliable biomarkers for Hepatocellular carcinoma(HCC). However, the underlying mechanism of AFP expression in HCC is poorly understood. In this study, we found that TCP10L, a gene specifically expressed in the liver, is down-regulated in HCC and that its expression inversely correlates with AFP expression. Moreover, overexpression of TCP10L suppresses AFP expression whereas knockdown of TCP10L increases AFP expression, suggesting that TCP10L might be a negative regulator of AFP. We found that TCP10L is associated with the AFP promoter and inhibits AFP promoter-driven transcriptional activity. Taken together, these results indicate that TCP10L negatively regulates AFP expression in HCC and that it could be a potential prognostic marker and therapeutic target for HCC.

계배 발생에 따른 닭 alpha-Fetoprotein의 분석

  • 박대규;유정아
    • 한국동물학회지
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    • 제32권4호
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    • pp.314-321
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    • 1989
  • 배의 발생과 분화에 따른 혈청 alpha-fetoprotein (AFP)의 생리적 기능 규명을 위한 기초실험의 일환으로, 모체와는 완전히 독립된 상태에서 발생과 분화가 이루어지는 계배의 혈청을 재료로 하여 CM-Sephadex C-50, hydroxyapatite 및 DEAE-Sepharose A-25등의 크로마토그라피 방법에 의해 닭 혈청 AFP를 분리하였다. 계 AFP는 분자량이 약 60 Kd로 사람 및 다른 포유동물 AFP의 분자량 (64-74 Kd)보다 작았다. 닭 AFP, 계배혈청 및 닭 혈청 등을 토끼에 면역하여 항체를 제조하였고, 이를 이용하여 닭의 배발생 단계에 따른 혈청 AFP 함량의 변화를 전기영동 및 면역학적 방법으로 정성 및 정량 분석을 실시하였다. 혈청 AFP 함량은 발생 7일-배 (AFP 농도 : 0.81 mg/ml)부터 점차 증가하여 13일-배에서 최고치인 2.46 mgfml로 나타났으며 그 이후 급격히 감소되어 부화 직후에는 매우 낮은 농도 (0.22mg/ml)였고 부화 4일에 거의 없어 졌으며, 이와 같은 AFP 함량의 변화는 혈청 알부민 함량의 변화와 거의 반비례 함을 알았다. For the preliminary step to investigate the site of AFP synthesis, the ontogenic characteristics and the physiological function of AFP, alpha-fetoprotein was isolated from chick embryo serum through the procedures of CM-Sephadex C-50, hvdroxvapatite and DEAE-Sephadex A-50 chro-matography. The isolated AFP was proved to be pure and its molecular weight was found to be about 60 Kd. With this purified AFP, rabbit anti-chick hor was produced. The serum AFP level in chick embryo has been investagated by using polyacrylamide gel electrophoresis, immunoelec-trophoresis and single radial immunodiffusion from 7 days of incubation until 4 days after hatch-ing. It was found that the AFP level is increased from 0.81 mg/ml at daw-7 embryo to maximum value of 2.46 mg/ml at day-13 embryo, followed by rapid decreases until hatching.

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황련추출물이 $CCl_4$중독 Rat의 간기능 회복에 미치는 영향 (Effects of Hwangryeun Extract on the Recovery of Liver Function in $CCl_4$-exposed Rats)

  • 김병우
    • 동의생리병리학회지
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    • 제25권6호
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    • pp.1020-1025
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    • 2011
  • Effects of Hwangryeun(Copitidis rhizoma) extract on recovery of liver function were researched in $CCl_4$ intoxicated rats. Concentration ${\alpha}$-fetoprotein, total protein in plasma showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups while that of albumin showed a increase. Activities of plasma aspartate aminotransferase and alanine aminotransferase in Hwangryeun(Copitidis rhizoma) extract applied groups showed a lower value than those of controlled groups. Alkaline phosphatase, lactate dehydrogenase and ${\gamma}$-glutamyl transpeptidase activities showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups. Concentration of plasma triglyceride and total cholesterol showed a lower values than that of control group.

Development of a One-step Two-site Enzyme Immunoassay for Measuring Human Alpha-fetoprotein by Eliminating Hook-effect

  • Kim, Se-Ho
    • BMB Reports
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    • 제34권1호
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    • pp.47-50
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    • 2001
  • A one-step, two-site enzyme immunoassay was developed for measuring human alpha-fetoprotein (AFP) in serum and amniotic fluid using monoclonal antibodies (McAb) by eliminating the high-dose hook effect. Three McAbs that recognize different epitopes were selected among 16 different clones on the basis of epitope mapping, two for immobilization and one for horseradish peroxidase conjugation. This one-step immunoassay system is more convenient and rapid compared to a conventional two-step sandwich immunoassay system. It did not exhibit the hook effect to around 2.7 mg/ml of AFP, which is probably one of the highest concentrations of AFP in the serum. The dose-response curve of the system was linear to 500 mg/ml of AFP and the system could differentiate as low as 1 mg/ml of AFP The intra- and inter-assay variations were in an acceptable range; 95~104% and 97~105% respectively Its correlation with other commercial systems was around 95%.

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A Rapid and Sensitive Two-Site Sandwich Enzyme-Linked Immunosorbent Assay for Detection of ${\alpha}$-Fetoprotein in Human Serum

  • Jang, Jeong-Su;Kim, Jeong-Min;Chung, Gi-Hyun;Paik, Bo-Hyun;Kim, Hack-Joo
    • 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.

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