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Kim, Sung-Hee,Cha, Sang-Ho,Karyn, Bischoff,Park, Sung-Won,Son, Seong-Wan,Kang, Hwan-Goo Korean Society of ToxicologyKorea Environmental Mu 2011 Toxicological Research Vol.28 No.2
Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.
Sung-Hee Kim,Sang-Ho Cha,Bischoff Karyn,Sung-Won Park,Seong-Wan Son,Hwan-Goo Kang 한국독성학회 2011 Toxicological Research Vol.27 No.2
Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with λ-type light chains. The IC??s of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/㎖, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/㎖, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/㎖ (R² > 0.99) and from 1 to 100 ng/㎖ (R² > 0.99), respectively. The intra- and interassay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.
Sang-Ho Cha,Sung-Hee Kim,Karyn Bischoff,Hyun-Jeong Kim,손성완,강환구 대한수의학회 2012 Journal of Veterinary Science Vol.13 No.2
A monoclonal antibody (mAb) against zearalenone (ZEN) was produced using ZEN-carboxymethoxylamine and -BSA conjugates. Antibody produced by one clone showing a very high binding ability was selected and found to have a higher affinity for ZEN compared to a commerciall ZEN antibody. We developed two direct competitive ELISA systems using the selected antibody (ZEN-coated and anti-ZEN antibody-coated ELISA). Quantitative ranges for the anti-ZEN antibody-coated ELISA and ZEN-coated ELISA were from 25 to 750 ppb and from 12.5 to 100 ppb, respectively. The detection limit of both methods as measured with standard solutions was 10 ppb. The intra-plate and inter-well variation of both ELISAs were less than 10%. The IC50 values for α -zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol compared to ZEN were 108.1, 119.3, 114.1, and 130.3% for the ZEN-coated ELISA. These values were 100.7, 120.7, 121.6, and 151.6% for the anti-ZEN antibody-coated ELISA. According to the anti-ZEN antibodycoated ELISA, the average recovery rates of ZEN from spiked animal feed containing 150 to 600 ng/mL of ZEN ranged from 106.07 to 123.00% with 0.93 to 2.28% coefficients of variation. Our results demonstrate that the mAb developed in this study could be used to simultaneously screen for ZEN and its metabolites in feed.
Provencal, J. L.,Montgomery, M. H.,Kanaan, A.,Thompson, S. E.,Dalessio, J.,Shipman, H. L.,Childers, D.,Clemens, J. C.,Rosen, R.,Henrique, P.,Bischoff-Kim, A.,Strickland, W.,Chandler, D.,Walter, B.,Wat IOP Publishing 2012 The Astrophysical journal Vol.751 No.2
<P>We report on an analysis of 308.3 hr of high-speed photometry targeting the pulsating DA white dwarf EC14012-1446. The data were acquired with the Whole Earth Telescope during the 2008 international observing run XCOV26. The Fourier transform of the light curve contains 19 independent frequencies and numerous combination frequencies. The dominant peaks are 1633.907, 1887.404, and 2504.897 mu Hz. Our analysis of the combination amplitudes reveals that the parent frequencies are consistent with modes of spherical degree l = 1. The combination amplitudes also provide m identifications for the largest amplitude parent frequencies. Our seismology analysis, which includes 2004-2007 archival data, confirms these identifications, provides constraints on additional frequencies, and finds an average period spacing of 41 s. Building on this foundation, we present nonlinear fits to high signal-to-noise light curves from the SOAR 4.1 m, McDonald 2.1 m, and KPNO 2 m telescopes. The fits indicate a time-averaged convective response timescale of tau(0) = 99.4 +/- 17 s, a temperature exponent N = 85 +/- 6.2, and an inclination angle of theta(i) = 32 degrees.9 +/- 3 degrees.2. We present our current empirical map of the convective response timescale across the DA instability strip.</P>