• Title/Summary/Keyword: serum antibody

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Comparative Analysis of Screening Results from Various ELISA Formats Used for Detection of Anti-Erythropoietin Antibodies in Korean Patients

  • Ha, Sung-Kyu;Yang, Seung-Ju;Shin, Sug-Kyun;Jo, Young-Il;Baek, Kyung-Min;Hong, Seung-Hwa;Pack, Seung-Pil;Kim, Sung-Jo;Heo, Tae-Hwe
    • Biomolecules & Therapeutics
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    • v.18 no.2
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    • pp.184-190
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    • 2010
  • Clinical cases of pure red cell aplasia (PRCA) have been reported during the recombinant human erythropoietin (EPO) therapy for the anemia patients. PRCA is a rare hematological disorder leading to a severe anemia due to an almost complete stop of red blood cell production. Antibody (Ab)-associated PRCA is caused by the EPO-neutralizing Abs that eliminate the biological activity of EPO. In order to detect anti-EPO Abs in human sera, we performed conventional ELISA, directly coated bridging ELISA, and streptavidin coated bridging ELISA, and compared their sensitivity and specificity. Some false positive results were obtained in the conventional ELISA. One positive sample was detected successfully by streptavidin coated bridging ELISA, which was not appeared in the directly coated bridging ELISA. In conclusion, streptavidin coated bridging ELISA was substantially sensitive and specific format and one out of sixty-eight serum samples was proved to be anti-EPO positive.

Comparison of chemiluminescence immunoassay (CIA) and immunochromatography assay(ICA) for Detecting HBsAg and Anti-HBs (B형 간염항원 및 항체 검사를 위한 화학발광면역검사법(CIA)과 면역크로마토그래피법(ICA)의 비교분석)

  • Kim, Hye-Jeong;Kim, Eun-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3419-3424
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    • 2013
  • Diverse immunoassays including a chemiluminescent immunoassay (CIA) are used to detect hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). Recently, an increasing number of institutions have been utilizing an immunochromatography assay (ICA), which is easy to use. In this study, We evaluated ICA kits for the rapid detection of HBsAg and anti-HBs by comparing them with a CIA. A total of 120 serum hospital samples, were collected for the whole month, were assayed using ICA kit. The Concordance rate, sensitivity, specificity, positive predictive value and negative predictive value of the ICA for HBsAg based on CIA results were 97%, 97%, 100%, 100%, and 96.8%, respectively. The diagnostic performances of the ICA for Anti-HBs were 90%, 90%, 93.3%, 93.1%, and 90.3%, respectively. The ICA kit failed to detect HBsAg and anti-HBs in low reactive samples. The ICA kits for the rapid detection of HBsAg might be recommended for interpreted with caution and dual analysis in the clinical laboratory.

Microplate Enzyme-Linked Immunosorbent Assay for Bovine Virus Antibody (우백혈병(牛白血病) Virus 항체측정(抗体測定)을 위한 효소면역법(酵素免疫法))

  • Choi, Won Pil
    • Current Research on Agriculture and Life Sciences
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    • v.1
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    • pp.195-199
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    • 1983
  • A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to bovine leukemia virus(BLV) is described and compared its sensitivity with that of the agar gel immunodiffusion test (ID) with BLV glycoprotein (gp) antigen using 263 sera collected in Korea and Japan. There was 98.5 per cent agreement between ELISA and ID when ELISA value, the value of tested serum(T) was devided with that of standard negative seurm(N) after the value of control was eliminated from T and N (T-C/N-C), of 1.5 or greater was considered positive. One hundred and forty four (99.6%) of 145 sera which were positive by ID were greater than 1.5 by ELISA, and 115 (97.5%) of 118 sera which were negative by ID were less than 1.5 by ELISA. As a result, it suggest that the ELISA test using BLV-gp antigen provides a useful serological tool for the diagnosis of BLV infection.

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Early Growth Response of Broilers to Dietary Lysine at Fixed Ratio to Crude Protein and Essential Amino Acids

  • Panda, A.K.;Rao, S.V. Rama;Raju, M.V.L.N.;Lavanya, G.;Reddy, E. Pradeep Kumar;Sunder, G. Shyam
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1623-1628
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    • 2011
  • The effects of dietary lysine (Lys) at a fixed ratio to crude protein (CP) and essential amino acids (EAA) on early growth response of broilers were studied. Four diets were formulated to contain similar metabolizable energy (ME, 2,950 kcal/kg) but contained graded levels of incremental Lys (1.1, 1.2, 1.3 and 1.4%) while also increasing the dietary CP and EAA (methionine, methionine+ cystine, threonine and tryptophan) to maintain a constant ratio with Lys. Each diet was fed at random to 10 replicates of 6 chicks each throughout the experimental period (1-21 d). At the lowest concentration of Lys of 1.1% (19.04% CP), body weight gain (BWG) was lowest and feed conversion ratio (FCR) was poorest. The BWG increased and FCR decreased linearly as dietary Lys increased upto 1.3% (22.5% CP). Lowest feed consumption was observed in the dietary group that contained 1.1% Lys (19.04% CP) in the diet. Increasing the concentration of Lys to 1.2% (20.77% CP), significantly increased the feed consumption. The concentrations of protein, calcium, phosphorus and cholesterol in serum were not influenced by the variation in Lys contents in the diet. The humoral immune response as measured by antibody titre in response to SRBC inoculation was significantly lower in the diets containing 1.1% Lys compared to 1.4%. It is concluded that the Lys requirement of broilers is 1.3% (22.5% CP) during 0 to 21 days of age for eliciting optimum performance when a fixed ratio of Lys to CP (1:17.31) and essential AA is maintained (1:0.47 Met; 1:0.56 Thr; 1:0.17 Try).

Recombinant DNA and Protein Vaccines for Foot-and-mouth Disease Induce Humoral and Cellular Immune Responses in Mice

  • Bae, Ji-Young;Moon, Sun-Hwa;Choi, Jung-Ah;Park, Jong-Sug;Hahn, Bum-Soo;Kim, Ki-Yong;Kim, Byung-Han;Song, Jae-Young;Kwon, Dae-Hyuck;Lee, Suk-Chan;Kim, Jong-Bum;Yang, Joo-Sung
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.265-273
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    • 2009
  • Foot-and-mouth disease virus (FMDV) is a small single-stranded RNA virus which belongs to the family Picornaviridae, genus Apthovirus. It is a principal cause of FMD which is highly contagious in livestock. In a wild type virus infection, infected animals usually elicit antibodies against structural and non-structural protein of FMDV. A structural protein, VP1, is involved in neutralization of virus particle, and has both B and T cell epitopes. A RNA-dependent RNA polymerase, 3D, is highly conserved among other serotypes and strongly immunogenic, therefore, we selected VP1 and 3D as vaccine targets. VP1 and 3D genes were codon-optimized to enhance protein expression level and cloned into mammalian expression vector. To produce recombinant protein, VP1 and 3D genes were also cloned into pET vector. The VP1 and 3D DNA or proteins were co-immunized into 5 weeks old BALB/C mice. Antigen-specific serum antibody (Ab) responses were detected by Ab ELISA. Cellular immune response against VP1 and 3D was confirmed by ELISpot assay. The results showed that all DNA- and protein-immunized groups induced cellular immune responses, suggesting that both DNA and recombinant protein vaccine administration efficiently induced Ag-specific humoral and cellular immune responses.

A Hospital-Based Serological Survey of Cryptosporidiosis in the Republic of Korea

  • Lee, Jong-Kyu;Han, Eun-Taek;Huh, Sun;Park, Woo-Yoon;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
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    • v.47 no.3
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    • pp.219-225
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    • 2009
  • The seroprevalence of cryptosporidiosis was examined using patients' sera collected from hospitals located in 4 different areas of the Republic of Korea. ELISA was used to measure antibody titers against Cryptosporidium parvum antigens from a total of 2,394 serum samples, which were collected randomly from patients in local hospitals; 1)Chungbuk National University Hospital, 2) Konkuk University Hospital, 3) local hospitals in Chuncheon, Gangwon-do (province), 4) Jeonnam National University Hospital, from 2002 through 2003. Of the2,394 samples assayed, 34%, 26%, and 56% were positive for C. parvum-specific IgG, IgM, and IgA antibodies, respectively, Positive IgG titers were most common in sera from Jeonnam National University Hospital, Gwangju, Jeollanam-do, and positive IgM titers were most common in sera from Chungbuk National University Hospital, Cheongju, Chuncheonqbuk-do. The seropositivity was positively correlated with age for both the IgG and IgA antibodies but was negatively correlated with age for the IgM antibodies. Western blotting revealed that 92%, 83%, and 77% of sera positive for IgG, IgM, and IgA ELISA reacted with 27-kDa antigens, respectively. These results suggested that infection with Cryptosporidium in hospital patients occurs more commonly than previously reported in the Republic of Korea.

Evaluation of Sperm Fertilizing Capacity Using Acrobeads Test (Acrobeads Test를 이용한 정자수정능의 평가)

  • Park, Young-Soo;Park, Nam-Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.81-86
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    • 1996
  • The assessment of acrosomal status is important in evaluating the ability of sperm to fertilize the egg. The acrosomal status of sperm from 47 normal volunteers with proven fertility and 167 subfertile men with not to achieve pregnancy for at least 1 year were evaluated with Acrobeads test(FUSO Pharmaceutical Industries, Ltd, Japan) using immunobeads coated with MH61 monoclonal antibody, which is specific for acrosome-reacted sperm. The mean${\pm}$SD of acrobeads score in 47 volunteer group was $2.8{\pm}0.7$, of which 46(97.9%)cases were ${\geq}$ 2. The mean${\pm}$SD of acrobeads score in 167 subfertile group was $1.7{\pm}0.8$, of which 73(79.3%)cases were ${\leq}$ 1. The aerobe ads score in subfertile group were significantly lower(r=0.294, p<0.05) than those in volunteer group. In subfertile group, acrobeads score were well correlated with the sperm density and motility(r=0.275, r=0.281, p<0.01), but not with semen volume(r=0.16) and serum hormone level(FSH r=0.084, LH r=0.036, testosterone r=0.058, prolactin r=0.006 and estradiol r=0.060)(p>0.05). Of 63 subfertile cases with normozoospermia, 22(34.9%)cases showed 0 or 1 of acrobeads score, which means to accompany with a functional defect in spite of normal morphology. As a results, Acrobeads test is not only a technically simple sensitive procedure with good reproducibility in evaluating the sperm fertilizing capacity but also an useful in the evaluation of effectiveness in the treatment of infertility and the separation of acrosome-reacted sperm in the assisted reproductive technique.

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Kinetics of 125I-BSA Binding to Monoclonal Anti-BSA Immobilized on $0.5{\mu}m$ Polystyrene Beads (c크기의 폴리스타이렌 담체에 고정화된 단세포군 항체와 항원 BSA의 속도론)

  • ;Dan
    • KSBB Journal
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    • v.11 no.2
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    • pp.225-237
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    • 1996
  • Intrinsic binding kinetics of of 125I-bovine serum albumin (125I-BSA) and immobilized monoclonal anti-BSA (MAb 9.1) were studied. Small non-porous polystyrene beads (0.5${\mu}$m diameter) were used as a solid support to minimize the mass transfer interference on rate measurements. We demonstrated both theoretically and experimentally that the binding reaction is kinetically controlled. Rate measurements show that the association reaction is of second order and the dissociation reaction is of first order. Between 4 and $37^{\circ}C$ the measured equilibrium constant agrees well with the equilibrium constant calculated from the rate measurements. The temperature effects on association are much greater than on dissociation; the activation energy for association is about 9Kca1/mole, as compared to 2Kca1/mole for dissociation. The use of small non-porous beads as a solid support in binding studies essentially avoids mass transfer limitations; such a system makes it possible to determine the intrinsic binding characteristics of any immobilized antibody on a solid surface.

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Fortification of Amino Acids to Improve Hybridoma Cell Growth and Monoclonal Antibody Production in Perfusion Culture (Perfusion배양시 세포성장 및 항체생산 향상을 위한 아미노산의 보강)

  • 이수영;최병욱;오한규;윤정원;전복환;변태호;박송용
    • KSBB Journal
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    • v.14 no.2
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    • pp.188-191
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    • 1999
  • We have investigated the fortifying effect of amino acids on the cell growth and productivity during the perfusion culture of hybridoma vR8 cells in serum-free media. Through the quantitative analysis of amino acids and metabolites in perfusion culture, we found that many amino acids(glutamine, histidine, arginine, methionine, isoleucine, leucine, phenylalanine, tryptophane) were heavily consumed at cell density of $1.06{\times}10^7$cells/mL. Due to amino acid depletion, cells died suddenly. So we supplemented the media with those amino acids by 30-170%. As a result, were could increase maximum cell density by 270%, average specific productivity by 175%, and average volumetric productivity by 560% in this fortified media, GC-HY-S2.

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A Study on the distribution of Anti-OEP, protease and elastase HA titer of Pseudomonas aeruginosa in healthy mother and infant (한국 건강유아(健康乳兒) 및 모체(母體)에 있어서 녹농균(綠膿菌)의 OEP, protease 및 elastase에 대(對)한 혈구응집가(血球凝集價))

  • Kim, Hae-Keun;Cho, Yang-Ja
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.39-46
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    • 1980
  • This experiment has been made to evaluate the distribution of OEP-HA titer, protease-HA titer and elastase-HA titer of antibody which are the common antigen of Pseudomonas aeruginosa in the serum of healthy mother-newborn infant-pairs, term pregnancy women and under 7 month old infants. By analysing the normal limt of these antibodies, it is expected that the result can be clinically applicable to the diagnosis, management and prevention of Pseudomonas aeruginosa infection, which shows rapid increase nowadays, 1. The Anti OEP-HA titer showed under 1:8, a very low titer, in 73% of cord sera group, but over 1:32 in 68% of mother sera group. 2. In healthy mother-newborn infant-pairs group, the anti OEP-HA titer of mothers showed 1:56, 48, much higher than that of newborn infant, 1:16.42. from which it can be concluded that their titer has no relation between the two. 3. The anti OEP-HA titers of healthy mother and term pregnancy women showed 1:56.5 and 1:53, 4, respectively, which are very similar to each ether, but in infant group, the titer showed 1:33, 51 higher than that of cord sera, 1:16.42 4. The anti protease-HA titer showed under 1:8 in 64-77.5% of total cases, which is lower than that of anti OEP-HA titer. 5. The anti elastase-HA titer showed under 1:8 in 93.75% of cord sera group and in 70.27-76.37% of infant, term perenancy women and mother group, but in infant group, the anti elastase-HA titer showed 1:16.56 higher than that of cord sera group 1:8.5.

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