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Development of a fluorescent microplate assay for determining cyanovirin-N levels in plasma

  1. Author:
    Bringans, S. D.
    O'Keefe, B. R.
    Bray, M.
    Whitehouse, C. A.
    Boyd, M. R.

  2. Author Address

    NCI, Mol Targets Dev Program, Ctr Canc Res, Frederick, MD 21702 USA. USA, Med Res Inst Infect Dis, Div Virol, Frederick, MD 21702 USA. Univ S Alabama, Coll Med, USA Canc Res Inst, Mobile, AL 36688 USA O'Keefe, BR, NCI, Mol Targets Dev Program, Ctr Canc Res, Bldg 562,Room 201, Frederick, MD 21702 USA
    1. Year: 2004
    2. Date: SEP
  2. Journal: Analytical and Bioanalytical Chemistry
    1. 380
    2. 2
    3. Pages: 269-274
  4. Type of Article: Article
  1. Abstract:

    A sensitive immunosorbent competition assay was developed for quantitation of the anti-HIV protein cyanovirin-N (CV-N) in plasma using a 96-well plate format and a fluorescent endpoint. The assay is based on the binding of CV-N in plasma to plate-bound anti-CVN antibodies, followed by removal of the plasma and addition of europium-labeled CV-N (Eu3+-CV-N) to compete for the remaining antibody sites. Detection by addition of a dissociative fluorescence enhancement solution and time-resolved fluorescence measurements allowed correlation to the concentration of the native CV-N in plasma. A linear detection range of 1-100 nM (r(2)>0.99) was obtained for CV-N in mouse plasma. This assay was then utilized for analysis of plasma levels of CV-N samples following subcutaneous injection of CV-N into mice. The results of these studies confirmed the reliability and sensitivity of this assay and the feasibility of its use for pharmacokinetic studies in a variety of species

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  1. View record in Web of Science®
  2. WOS: 000224528900013

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