• 제목/요약/키워드: Antibody screening

검색결과 156건 처리시간 0.026초

Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • 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.

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Antibody-based Screening of Porphyromonas gingivalis Proteins Specifically Produced in Patients with Chronic Periodontitis

  • Kim, Hye-Jung;Lee, Seok-Woo
    • International Journal of Oral Biology
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    • 제43권4호
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    • pp.201-207
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    • 2018
  • Porphyromonas gingivalis is among the major etiological pathogens of chronic periodontitis. The virulence mechanisms of P. gingivalis is yet to be identified as its activity is largely unknown in actual disease process. The purpose of this study is to identify antigens of P. gingivalis expressed only in patients with chronic periodontitis using a unique immunoscreening technique. Change Mediated Antigen Technology (CMAT), an antibody-based screening technique, was used to identify virulence-associated proteins of P. gingivalis that are expressed only during infection stage in patients having chronic periodontitis. Out of 13,000 recombinant clones screened, 22 tested positive for reproducible reactivity with rabbit hyperimmune anti-sera prepared against dental plaque samples acquired from periodontitis patients. The DNA sequences of these 18 genes were determined. CMAT-identified protein antigens of P. gingivalis included proteins involved in energy metabolism and biosynthesis, heme and iron binding, drug resistance, specific enzyme activities, and unknown functions. Further analysis of these genes could result in a novel insight into the virulence mechanisms of P. gingivalis.

대전지역 일개 종합병원에서의 비예기 항체 발생현황과 분포 (Frequencies and Distributions of Unexpected Antibodies at a General Hospital in the Daejeon of Korea)

  • 김재중
    • 대한임상검사과학회지
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    • 제50권3호
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    • pp.354-358
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    • 2018
  • 수혈 준비 시 비예기 항체 선별검사를 통한 항체의 유무와 그 종류를 아는 것은 매우 중요하다. DiaMed-ID시스템을 이용하여 대전지역 일개 대학병원에 2016년 1월부터 2017년 12월까지 2년 동안 의뢰된 비예기 항체 선별검사 양성자 중에서 항체동정은 55명에서만 되었고 주로 여성에서 자주 동정되었다. Rh 36예(65.5%), Lewis 7예(12.7%), Kidd 4예(7.3%), Duffy 4예(7.3%), MNSs 3예(5.5%), Rh+Kidd 복합 1예(1.8%)가 동정되었는데, Rh 계열은 Anti-E 19예(34.5%), Anti-E/-c 4예(7.3%), Anti-C/-e 4예(7.3%), $Anti-E/-c/-Jk^b$ 1예(1.8%)가, Lewis 계열은 $Anti-Le^a$ 3예(5.5%), $Anti-Le^b$ 3예(5.5%), Kidd 계열은 $Anti-Jk^a$ 1예(1.8%), $Anti-Jk^b$ 3예(5.5%)가 동정되었다. Duffy와 MNSs 계열은 각각 $Anti-Fy^a$ 1예(1.8%), $Anti-Fy^b$ 3예(5.5%), Anti-M 2예(3.6%), Anti-S 1예(1.8%)가 동정되었다. 최근 대전의 비예기 항체 빈도와 분포가 반영된 이 연구는 효율적인 수혈준비에 도움이 될 것이다.

잔류 Chloramphenicol 검사용 효소 면역측정법의 개발에 관한 연구 (Study on Development of Enzyme-Linked Immunosorbent Assay for the Screening of Chloramphenicol Residues)

  • 윤동호;이문한
    • 한국식품위생안전성학회지
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    • 제8권4호
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    • pp.205-214
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    • 1993
  • The monoclonal antibody to chloramphenicol(CAP) was produced to develop an enzyme-linked immunosorbent assay(ELISA) for residual CAP. An immunogen(CAP-BSA) was prepared by immunogen, antibody titer was measured by indirect ELISA. Spleen cells form the immunized mouse were fused with SP2/OAg14 myeloma cells. Among hybridomas selected in HAT media, 6 clones shown high antibody titer to CAP were subjected to cloning by limit dilution, and all of the monoclonal antibodies(MCA1, 2, 3, 4, 5, 7 and 9) produced by each clone were identified as IgG1 by ELISA isotyping analysis. Competitive ELISA condition was established by using the purified monoclonal antibody MCA1 as primary antibody and CAP-HSA conjugate as coating antigen. Standard curve of CAP(n=28) showed that the lowest detection limit of CAP is 20ng/ml level. The cross-reactivities of the 6 monoclonal antibodies showed that CAP sodium succinate. CAP base, P-nitrophenol, and p-nitrobenzyl alcohol were 89∼178, 0.050∼2.237, 0.056∼0.794 and 0.013∼7.939%, respectively. No cross-reactivities were observed with phenylalanine, tyrosine, glutamine, thiamphenicol, neomycin, streptomycin, gentamicin, sulfamethazine, sulfathiazole, chlortetracycline and p-aminobenzoic acid.

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Confidence intervals for the COVID-19 neutralizing antibody retention rate in the Korean population

  • Apio, Catherine;Kamruzzaman, Md.;Park, Taesung
    • Genomics & Informatics
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    • 제18권3호
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    • pp.31.1-31.8
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    • 2020
  • The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global pandemic. No specific therapeutic agents or vaccines for COVID-19 are available, though several antiviral drugs, are under investigation as treatment agents for COVID-19. The use of convalescent plasma transfusion that contain neutralizing antibodies for COVID-19 has become the major focus. This requires mass screening of populations for these antibodies. While several countries started reporting population based antibody rate, its simple point estimate may be misinterpreted without proper estimation of standard error and confidence intervals. In this paper, we review the importance of antibody studies and present the 95% confidence intervals COVID-19 antibody rate for the Korean population using two recently performed antibody tests in Korea. Due to the sparsity of data, the estimation of confidence interval is a big challenge. Thus, we consider several confidence intervals using Asymptotic, Exact and Bayesian estimation methods. In this article, we found that the Wald method gives the narrowest interval among all Asymptotic methods whereas mid p-value gives the narrowest among all Exact methods and Jeffrey's method gives the narrowest from Bayesian method. The most conservative 95% confidence interval estimation shows that as of 00:00 on September 15, 2020, at least 32,602 people were infected but not confirmed in Korea.

Identification of HUGT1 as a Potential BiP Activator and a Cellular Target for Improvement of Recombinant Protein Production Using a cDNA Screening System

  • Ku, Sebastian Chih Yuan;Lwa, Teng Rhui;Giam, Maybelline;Yap, Miranda Gek Sim;Chao, Sheng-Hao
    • Molecules and Cells
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    • 제27권5호
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    • pp.577-582
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    • 2009
  • The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ${\gamma}$, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.

조기 난소기능 부전증 환자에서 자가면역 질환과의 상관관계 (Association with Autoimmune Disease in Patients with Premature Ovarian Failure)

  • 박준철;김종인;이정호
    • Clinical and Experimental Reproductive Medicine
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    • 제31권3호
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    • pp.149-154
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    • 2004
  • Objective: To assess the association with autoimmune endocrine diseases and detection rate of autoimmune antibodies and its clinical significance in patients with premature ovarian failure. Methods: Twenty eight patients with primary or secondary amenorrhea manifesting hormonal and clinical features of premature ovarian failure (primary POF: 7, secondary POF: 21) were investigated. We tested them TFT, 75 g OGTT, ACTH and S-cortisol for thyroiditis, IDDM, Addison's disease, and antithyoglobulin antibody, antimicrosomal antibody, antinuclear antibody, rheumatic factor, anti-smooth muscle antibody, anti-acetylcholine receptor antibody for non-organ specific autoimmune disorders. Results: Only one patient was diagnosed as IDDM and no patients had abnormal TFT or adrenal function test. More than one kind of autoantibody was detected in 11 patients of all (39.2%): 5 patients (71.4%) of primary POF group and 6 patients (21.4%) of secondary POF group. Eleven patients (39.3%) had antithyroglobulin antibody, 4 (14.3%) had antimicrosomal antibody, 2 (7.1%) had antinuclear antibody, 2 (7.1%) had rheumatic factor, 1 (3.6%) had anti-smooth muscle antibody, 1 (3.6%) had anti-acetylcholine receptor antibody. Conclusions: Premature ovarian failure may occur as a component of an autoimmune polyglandular syndrome, so patients should be measured with free thyroxine, thyroid-stimulating hormone, fasting glucose and electrolytes. Measurement of thyroid autoantibodies in POF patients may be important in identifying patients at risk of developing overt hypothyoidism, but other autoantibodies may not be suitable for screening test.

비예기항체 선별검사에서 항-Lua의 검출: 증례보고 및 문헌고찰 (Detection of Anti-Lua in an Unexpected Antibody Screening Test: A Case Report and Literature Review)

  • 송새암;오승환;박태성;손혜수;성성경;이자영;전경란;신정환;김혜란;이정녀
    • 대한수혈학회지
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    • 제23권2호
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    • pp.169-172
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    • 2012
  • Lutheran a ($Lu^a$) 항원은 백인과 흑인에서 6~8%의 낮은 빈도로 발견되며 한국인을 포함한 그 외의 인종에서는 거의 발현되지 않는다. 따라서 $Lu^a$가 상당한 면역원성을 가지고 있음에도 $Lu^a$에 감작되는 경우는 거의 없다. 본 증례에서는 Lu (a-/b+)형을 가진 70세 여자 환자에서 발생한 $Lu^a$에 대한 희귀한 항체를 보고하며 관련 문헌을 고찰하였다. 비예기항체 선별검사 및 동정검사에서 사용되는 패널 중 $Lu^a$ 양성 적혈구가 부족하여 이러한 드문 $Lu^a$에 대한 항체를 검출하는 것이 쉽지 않다. 따라서, 이러한 낮은 빈도의 항원에 대한 항체를 검출하는 데 있어서 위음성의 가능성을 항상 염두에 두어야 한다.

Development of Immunochromatography Strip-Test Using Nanocolloidal Gold-Antibody Probe for the Rapid Detection of Aflatoxin B1 in Grain and Feed Samples

  • Shim, Won-Bo;Yang, Zheng-You;Kim, Jung-Sook;Kim, Ji-Young;Kang, Sung-Jo;Woo, Gun-Jo;Chung, Young-Chul;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • 제17권10호
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    • pp.1629-1637
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    • 2007
  • An immunochromatography (ICG) strip test using a nanocolloidal gold-antibody probe was developed and optimized for the rapid detection of aflatoxin B1 (AFB1). A monoclonal antibody specific to AFB1 was produced from the cloned hybridoma cell (AF78), coupled with nanocolloidal gold, and distributed on the conjugate pad of the ICG strip test. The visual detection limit of the ICG strip test was 0.5 ng/ml, and this method showed a cross-reaction to aflatoxin B2, G1, and G2. In total, 172 grain and feed samples were collected and analyzed by both the ICG strip test and HPLC. The results of the ICG strip test showed a good agreement with those obtained by HPLC. These results indicated that the ICG strip test has a potential use as a rapid and cost-effective screening tool for the determination of AFB1 in real samples and could be applied to the preliminary screening of mycotoxin in food and agricultural products, generating results within 15 min without complicated steps.

Diagnostic Value of Immunoglobulin G Anti-Deamidated Gliadin Peptide Antibody for Diagnosis of Pediatric Celiac Disease: A Study from Shiraz, Iran

  • Anbardar, Mohammad Hossein;Haghighi, Fatemeh Golbon;Honar, Naser;Zahmatkeshan, Mozhgan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제25권4호
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    • pp.312-320
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    • 2022
  • Purpose: Screening serologic tests are important tools for the diagnosis of celiac disease (CD). Immunoglobulin (Ig)G anti-deamidated gliadin peptide (anti-DGP) is a relatively new autoantibody thought to have good diagnostic accuracy, comparable to that of anti-tissue transglutaminase (anti-tTG) antibody. Methods: Pediatric patients (n=86) with a clinical suspicion of CD were included. Duodenal biopsy, anti-tTG, and IgG anti-DGP antibody tests were performed. The patients were divided into CD and control groups based on the pathological evaluation of duodenal biopsies. The diagnostic accuracy of serological tests was determined. Results: IgA anti-tTG and IgG anti-DGP antibodies were positive in 86.3% and 95.4% of patients, respectively. The sensitivity, specificity, and diagnostic accuracy of the IgA anti-tTG test were 86.3%, 50.0%, and 68.6%, respectively, and those of the IgG anti-DGP test were 95.4%, 85.7%, and 90.7%, respectively. The area under the receiver operating characteristic (ROC) curve was 0.84 (95% confidence interval [CI], 0.74-0.91) for IgA anti-tTG test and 0.93 (95% CI, 0.86-0.97) for IgG anti-DGP test. The comparison of IgA anti-tTG and IgG anti-DGP ROC curves showed a higher sensitivity and specificity of the IgG anti-DGP test. Conclusion: IgG anti-DGP is a reliable serological test for CD diagnosis in children. High tTG and DGP titers in the serum are suggestive of severe duodenal atrophy. The combined use of IgA anti-tTG and IgG anti-DGP tests for the initial screening of CD can improve diagnostic sensitivity.