Comparative sensitivity of immunoassays for haptens using monomeric and dimeric antibody fragments

被引：22
|
作者：
Grant, SD [1 ]
Porter, AJ [1 ]
Harris, WJ [1 ]
机构：
[1] Univ Aberdeen, Inst Med Sci, Dept Mol & Cell Biol, Aberdeen AB25 2ZD, Scotland
来源：
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 1999年 / 47卷 / 01期
关键词：
antibody fragments; immunoassays; single-chain antibodies; anti-haptens; triazines;
D O I：
10.1021/jf9808574
中图分类号：
S [农业科学];
学科分类号：
09 ;
摘要：
A single-chain anti-atrazine antibody fragment, scAb (single-chain Fv with a CK domain), was expressed in Escherichia coli, and monomeric and dimeric species were preferentially purified from periplasmic extracts by chromatography upon nickel chelate immunosorbent columns or by immunoaffinity purification using a constant domain (CK) tag. Recombinant monomeric and dimeric antibody fragments, Fab, and intact monoclonal antibodies were compared in assays by competition between free atrazine in solution and (a) immobilized atrazine-bovine serum albumin conjugate (indirect assay) or (b) atrazine-alkaline phosphatase (direct assay). Recombinant antibody fragments provided a lower detection Limit than either Fab or intact monoclonal antibody in both assay formats. Monomeric fragments displayed a sensitivity of detection down to 0.1 ppb, compared to 1.0 ppb for dimeric fragments and the parental monoclonal.
引用
收藏
页码：340 / 345
页数：6
相关论文
共 50 条
  • [31] Sensitivity Improvement in Electrochemical Immunoassays Using Antibody Immobilized Magnetic Nanoparticles with a Clean ITO Working Electrode
    Sunga Song
    Young Joo Kim
    Hye-Lim Kang
    Sumi Yoon
    Dong-Ki Hong
    Won-Hyo Kim
    Ik-Soo Shin
    Woo Kyeong Seong
    Kook-Nyung Lee
    BioChip Journal, 2020, 14 : 308 - 316
  • [32] Enhancing antibody detection sensitivity in lateral flow immunoassays using endospores of Bacillus subtilis as signal amplifiers
    Lin, Wen-Zhi
    Hung, Chin-Mao
    Lin, I-Hsien
    Sun, Yi-Jia
    Liao, Zheng-Xiu
    Wu, Chia-Chun
    Hou, Shao-Yi
    TALANTA, 2024, 276
  • [33] Structural and mutational analysis of a monomeric and dimeric form of a single domain antibody with implications for protein misfolding
    George, Jade
    Compton, Jaimee R.
    Leary, Dagmar H.
    Olson, Mark A.
    Legler, Patricia M.
    PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 2014, 82 (11) : 3101 - 3116
  • [34] Multiple antibody conjugation to PEG grafted copolymer improves sensitivity of immunoassays
    Trimaille, Thomas
    Verrier, Bernard
    Coiffier, Celine
    Gigmes, Didier
    EUROPEAN POLYMER JOURNAL, 2024, 216
  • [35] Designing novel nano-immunoassays: antibody orientation versus sensitivity
    Puertas, S.
    Moros, M.
    Fernandez-Pacheco, R.
    Ibarra, M. R.
    Grazu, V.
    de la Fuente, J. M.
    JOURNAL OF PHYSICS D-APPLIED PHYSICS, 2010, 43 (47)
  • [36] The importance of antibody orientation for enhancing sensitivity and selectivity in lateral flow immunoassays
    Lu, Zhao-Yu
    Chan, Yang-Hsiang
    SENSORS & DIAGNOSTICS, 2024, 3 (10): : 1613 - 1634
  • [37] Multiple antibody conjugation to PEG grafted copolymer improves sensitivity of immunoassays
    Trimaille, Thomas
    Verrier, Bernard
    Coiffier, Céline
    Gigmes, Didier
    European Polymer Journal, 2024, 216
  • [38] SENSITIVITY OF MONOMERIC AND DIMERIC FORMS OF ESCHERICHIA-COLI THYMIDINE KINASE TO ULTRAVIOLET-RADIATION
    CYSYK, RL
    PRUSOFF, WH
    PHOTOCHEMISTRY AND PHOTOBIOLOGY, 1972, 15 (06) : 589 - &
  • [39] PREPARATION AND BIOLOGIC CHARACTERIZATION OF FRAGMENTS CONTAINING DIMERIC AND MONOMERIC C-GAMMA-2 DOMAIN OF RABBIT IGG
    UTSUMI, S
    OKADA, M
    UDAKA, K
    AMANO, T
    MOLECULAR IMMUNOLOGY, 1985, 22 (07) : 811 - 819
  • [40] USE OF MONOCLONAL-ANTIBODY TO INCREASE THE SENSITIVITY AND SPECIFICITY OF PRECIPITATING IMMUNOASSAYS AND CELL-SURFACE BINDING IMMUNOASSAYS
    MONTGOMERY, RR
    KUNICKI, TJ
    GLODE, LM
    METHODS IN ENZYMOLOGY, 1986, 121 : 702 - 717
12345