• Title/Summary/Keyword: enzyme linked immuno sorbent assay(ELISA)

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Studies on the development of enzyme linked immuno-sorbent assay (ELISA) for hepatitis B surface antigen (HBsAg) by monoclonal antibodies of different affinity constants

  • Kim, Gye-Won;Hong, Sung-Youl;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.18-24
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    • 1987
  • Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

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Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins (우 Casein의 면역효소분석법)

  • 염행철
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.87-102
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    • 1992
  • A rapid, sensitive, and specific enzyme-linked immuno-sorbent assay (ELISA) for bovine casein was developed. Biotinylated casein and peroxidase-conjugated avidin were used in the assay with antibody separated from yolks of immunized hens. Caseins were biotinylated with sulfo-N-hydroxy succinimido biotin and peroxi-dase-conjugated avidin bound the biotinylated casein which became bound to immobilized anti-body on a microplate. The antibodies were specific for bovine $\alpha$- and $\beta$-caseins, and their cross-reactivities with whey proteins, IgG, and serum albumin from bovine were not detectable by ELISA and Western blot. Various sensitivities ranging from 2ng/ml to 20${\mu}\textrm{g}$/ml of casein were achieved, and were controlled by adding vanous concentrations of the biotinylated casein. Parallelism was observed between standard and sample curves. The coefficients of variation of intra-assays and inter-assays from the most sensitive assay were 5.5 and 5.7%, respectively, at the 50% displacement. Casein contents of peripaturient milk samples showed that casein secretion rapidly increased 3d prepartum.

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Enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus (축우 부루셀라병의 ELISA 진단법에 관한 연구)

  • Lim, Yoon-kyu;Lee, Doo-sick;Park, Jun-hong;Yang, Ki-chun;Kim, Seung-ho;Kim, Kong-sick;Hyun, Kwan-jong;Kim, Woo-tack;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.131-135
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    • 1993
  • Enzyme-linked Immuno sorbent Assay (ELISA) for the serological diagnosis of Brucella abortus was developed and compared with plate agglutination test. Cell wall antigen was extracted from Brucella abortus 1119-3 by sonication and with a sodium deoxychlate solution. Optimum protein concentration of coating antigen were $0.4{\mu}g/100{\mu}{\ell}$ protein on each microtiter plate well. Horse radish peroxidase (HRP) labeled protein-G was used as a tracer of reacted antibodies. ELISA confirmed the agreeable results of 40 cases out of 43 cases by plate aggulutination test. ELISA diagnosed positive cases(10 out of 12) and negative cases (1 out of 12) with dubious sera by plate agglutination test. From this results ELISA could be used for the early diagnostic tools of Brucellosis in cattle.

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Development and Characterization of Anti-gliadin Polyclonal Antibody in Wheat

  • Chang, Suk Joo;Hong, Byung Hee;Seo, Yang Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.4
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    • pp.339-344
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    • 1999
  • Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

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Development of a Quantitative ELISA for Anti HER-2 Antibodies using Human HER-2 Recombinant Proteins (인간 HER-2 재조합 단백질을 사용한 항 HER-2 항체 단백질의 ELISA 정량 방법 개발)

  • Jung, Sun-Ki;Ryu, Chang-Seon;Choung, Kyu-Jin;Song, Gyu-Yong;Kim, Sang-Kyum
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.16-21
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    • 2011
  • HER-2 (Human Epidermal Growth Factor Receptor-2) is a protein giving higher aggressiveness in human breast cancers. Trastuzumab is a monoclonal antibody that targets HER-2 and is known to extend survival across all stages of HER2-positive breast cancer. In this study, we attempted to development of a quantitative ELISA (Enzyme-Linked ImmunoSorbent Assay) for evaluating anti HER-2 antibodies using human HER-2 recombinant proteins to support antibody producing processes and pharmacokinetic studies. We established direct or indirect ELISA method for the trastuzumab-like protein combined human recombinant HER-2. The ELISA method will prove to be great value in quantitating anti-HER-2 antibodies levels for developing anticancer antibodies.

A Study on the Mating Types and Serotypes of Clinical Isolates of Cryptococcus neoformans and Production of Serodiagnostic Antigen and Antiserum for Cryptococcosis (우리나라 환자로부터 분리된 Cryptococcus neoformans의 균학적 특성과 혈청학적 진단용 항원 및 항체생산에 관한 연구)

  • Kim, Sang-Jae;Kim, Sin-Ok;Lee, Seung-Ho;Chong, Yon-Sop;Suk, Jong-Sung
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.127-131
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    • 1986
  • The mating types and serotypes of 10 clinical isolates of Cryptococcus neoformans have been investigated. Seven isolates were serotype A and three were serotype D and thus they fell in C. neoformans var. neoformans. Mating types of six isolates were found $\alpha$ and two were $\alpha$ but another two isolates were untypable. Enzyme linked immuno-sorbent assay(ELISA) using rabbit hyper-immune serum to cryptococcal polysaccharides was well adapted to the analysis of capsular polysaccharides in sera of the patients with cryptococcal meningitis.

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Diagnosis of Atopy in Dogs : A Review of the Need for Conceptual and Diagnostic Refinement I (개들의 아토피 진단 : 개념적 및 진단학적 순화의 필요성 검토 I)

  • Perkins Jacqueline
    • Journal of the korean veterinary medical association
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    • v.38 no.4
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    • pp.376-382
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    • 2002
  • 호주의 많은 소동물 임상수의사들이 최근에 관심을 갖고 있는 부분은 개의 아토피(Canine Atopy)에서의 알러지-특이성(Allergen-specific)IgE에 쓰이는 혈청 ELISA (Enzyme-Linked Immuno-Sorbent Assay, 효소결합면역흡착검사)테스트의 활용 가능성에 관한 것이다. 희망사항은 개 아토피의 진단과 치료에 있어서 도움이 되도록 접근할 수 있는 시험(Accessible test)일 것이다. 아토피(Atopy)는 진단과 치료를 실패시키는 질병이며, 새로운 ELISA라도 이 문제를 쉽게 해결하여 주지 못한다. 개의 아토피에서 알려진(Allergens)들을 분석하는 데 쓰이는 혈청 ELISA처럼 아주 정확하게 특성화되어져야 한다. 해당 개는 아토피성(Atopic)으로서 진단되어져야만 되고 그 다음에 치료에 지침이 되도록 개별 알러진 동정(Individual allergen identification)용 ELISA로 개의 혈액을 스크린하는데 적합하여야 한다. ELISA와 피내접종시험(Internal testing)방법은 상관도가 낮고 그렇기 때문에 어떤 시험이 더 신뢰할 수 있는 것일까? 이러한 사실은 논쟁의 여지가 있으며 이상의 2가지 시험들이 옳다면 각각의 상응하는 각각 다른 아형(Subtypes)의 아토피라고 볼 수있다. 아마도 알러진들은 경피적으로 흡수되어 아토피성피부염으로 되어진다. 그것은 인체에서 건초열(Hay fever)이나 천식(Asthma)과 같이 흡입에 의해 일으키는 유사한 증후군동종 (Syndrome akin)과 같은 접근의 초기양식(Primary mode of access)으로부터 생기는 다른 유사한 질병과정이 되어질 수도 있다. 이러한 2가지 증후군은 함께 병발하는 경우가 있다. 의학에서는 피부 반점시험방법의 예비조사연구(Preliminary research with skin patch testing in medicine)는 아토피성 피부염을 야기하는 알러진들의 경피흡수경로의 개념을 지지해준다. 흡인 알러진들을 사용한 개들에서의 도전 시험(Challenge tests)들은 거의 대부분이 비-피부학적 임상증상들을 발현하였다. 모든 증거가 분명히 나타나도록 미래의 조사연구를 인도해 주는 게 필요하며 개 아토피의 정의와 진단을 둘러싼 수수께끼를 해결하는데 도움이 될 것이다.

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Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang (청심연자탕(淸心蓮子湯)이 태음인(太陰人) 뇌경색증(腦硬塞症) 환자(患者)의 세포활성물질(細胞活性物質) 생성조절(生成調節)에 미치는 영향(影響))

  • Kim, Kyung-Yo;Noh, Hyun-Soo
    • Journal of Sasang Constitutional Medicine
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    • v.12 no.2
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    • pp.162-170
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    • 2000
  • Purpose : Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang Method : ELISA(enzyme-linked immuno-sorbent assay) Result : Chungsimyeunjatang(CYT) is a prescription for the cerebral infarction (CI) patients of Taeumin according to Sasang constitution philosophy. Taeumin patients with CI were treated with CYT during the acute stage. Clinical signs of CI disappeared markedly in about two to four weeks after oral administration of CYT in all patients. The mean interleukin (IL)-2 plasma levels were slightly lower in the patients with CI than in the normal groups, whereas the mean IL-4, IL-6 and IgE levels were significantly higher in the patients. There were no significant differences in $interferon-{\gamma}$ $(IFN-{\gamma})$ levels between the groups. Serum $IFN-{\gamma}$ and IL-2 levels derived from T helper (Th)1 cells were elevated significantly in the patients with CI by CYT administration. Significant reduced plasma levels of IL-4 and IL-6 derived from Th2 cells and IgE were observed in the patients treated with CYT. During the period of CYT administration, there were no other adverse effects. The data indicate that CYT has a good CI treatment effect, and that its action may be due to regulation of cytokine Production.

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Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.