• Title/Summary/Keyword: A-fetoprotein

Search Result 99, Processing Time 0.025 seconds

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

  • Michung Yoon
    • Biomedical Science Letters
    • /
    • v.3 no.2
    • /
    • pp.115-123
    • /
    • 1997
  • $\alpha$-Fetoprotein (AFP) has been a useful marker in screening and/or monitoring patients with hepatocellular carcinoma, gonadal germ cell tumor, gastric carcinoma and neural tube defects. In the present study, it was attempted to produce anti-human AFP polyclonal antibodies and to develop a competitive enzyme-linked immunosorbent assay (ELISA) for the measurement of AFP in human plasma and amniotic fluid. AFP was isolated from amniotic fluid using an isolation procedure consisting of affinity chromatography and preparative polyacrylamide gel electrophoresis. The antibody directed against AFP was raised in rabbits. Double immunodiffusion and Western blotting methods showed that the antiserum was highly specific, reacting with only AFP-containing samples. Standard curve was obtained by using purified AFP and specific antiserum. The assay sensitivity was 5ng/ml and the working range was 5~l,000ng/ml. The within-assay and between-assay coefficient of variance (CV) was 4.5% and 8.5%, respectively. These results indicate that the assay is valuable for the measurement of AFP and found to be simple, reproducible, and accurate.

  • PDF

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

  • Kang, Shin-Sung;Bang, Ok-Sun;Park, Tae-Kyu
    • The Korean Journal of Zoology
    • /
    • v.29 no.4
    • /
    • pp.283-293
    • /
    • 1986
  • For the preliminary step to make and characterize the monoclonal antibodies of human alpha-fetoprotein (AFP) was purified from 534g of human fetal tissues through the procedures of tissue extraction, DEAE-cellulose, concanavalin A-Sepharose, Cibacron Blue F3GA-agarose and immunoadsorbent affinity chromatography. The isolated AFP preparation showed a single band on polyacrylamide gel electrophoresis and a single precipitin are against rabbit anti-human cord serum and anti-human AFP on immunoelectrophoresis. Our AFP also displayed a single band on SDS-polyacrylamide gel electrophoresis. The recovery of AFP was 8.76mg total.

  • PDF

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

  • Kim, Yoohyun
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.40 no.2
    • /
    • pp.86-93
    • /
    • 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.

  • PDF

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

  • Won, Kyoung-Sook;Ryu, Jin-Sook;Moon, Dae-Hyuk;Lee, Hee-Kyung
    • The Korean Journal of Nuclear Medicine
    • /
    • v.34 no.4
    • /
    • pp.353-359
    • /
    • 2000
  • Purpose: Although ${\alpha}$-fetoprotein is one of the most commonly used tumor markers in Korea, most of the radioimmunoassay kits for ${\alpha}$-fetoprotein have been imported from foreign countries. The purpose of this study was to evaluate the performance of a recently developed domestic immunoradiometric kit for ${\alpha}$-fetoprotein (Riakey AFP IRMA $CT^R$, Sin-Jin Medics, Seoul, Korea). Materials and Methods: We evaluated intra- and inter-assay precision, recovery rate, parallelism, and sensitivity of serum ${\alpha}$-fetoprotein measurement using Riakey AFP IRMA $CT^R$ kit. The values of ${\alpha}$-fetoprotein measured by Riakey AFP IRMA $CT^R$ kit were compared with those measured by two foreign commercial kits (${\alpha}$-fetoproteina of Radim and ${\alpha}$-feto.riabead of Abbott). Results: Intra-assay coefficients of variation on three different levels were 5.3% for 18.9 ng/ml, 3.4% for 133 ng/ml and 1.6% for 330 ng/ml. Inter-assay coefficients of variation were 9.7% for 20.9 ng/ml, 3.2% for 137 ng/ml and 4.1% for 330 ng/ml respectively. Recovery rate tests on all three different levels showed within $100{\pm}10%$. Parallelism was also good and the sensitivity was 0.63 ng/ml. There was strong correlation between the measurement of ${\alpha}$-fetoprotein by Riakey AFP IRMA $CT^R$ and that by two foreign commercial kits(r=0.98). Conclusion: The first Korean domestic immunoradiometric kit for ${\alpha}$-fetoprotein, Riakey AFP IRMA $CT^R$, performed well for clinical use.

  • PDF

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

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
    • /
    • v.7 no.3
    • /
    • pp.103-110
    • /
    • 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.

  • PDF

TCP10L negatively regulates alpha-fetoprotein expression in hepatocellular carcinoma

  • Shen, Suqin;Feng, Huan;Liu, Longjiang;Su, Wei;Yu, Long;Wu, Jiaxue
    • BMB Reports
    • /
    • v.53 no.8
    • /
    • pp.431-436
    • /
    • 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
    • The Korean Journal of Zoology
    • /
    • v.32 no.4
    • /
    • pp.314-321
    • /
    • 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.

  • PDF

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

  • Kim, Byoung-Woo
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.25 no.6
    • /
    • pp.1020-1025
    • /
    • 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
    • /
    • v.34 no.1
    • /
    • pp.47-50
    • /
    • 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%.

  • PDF

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
    • /
    • v.29 no.3
    • /
    • pp.192-199
    • /
    • 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.

  • PDF