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New adjuvant design using layered double hydroxide for production of polyclonal antibodies in radioimmunoassay techniques

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Abstract

Various adjuvants have been used to enhance the immune response against specific antigens. So the objective of this work describes the immune stimulating activity of layered double hydroxide (LDH) particles incorporate with mineral oil as a new formulation of adjuvant as compared to known Freund’s adjuvant for production of alpha-fetoprotein polyclonal antibody (anti-AFP) for estimation of alpha fetoprotein (AFP) in human serum by radioimmunoassay technique. In this concern, the study comprised two groups of white New Zealand rabbits, 2−2.5 kg body weight and each group comprised three rabbits. The first group vaccinated with AFP antigen emulsified with Freund’s adjuvant and the second group vaccinated with AFP antigen emulsified with LDH formulation. The obtained data show that the highest displacement using LDH adjuvant reached (74.2, 61.7 and 66.5 %) while the corresponding values with Freund’s adjuvant recorded (64.8, 60.3 and 54.6 %) which indicates that the use of LDH adjuvant as a cellular vehicle is a more suitable choice. Also, the preparation of AFP tracer using lactoperoxidase oxidation method and its purification using gel chromatography on PD-10 column were carried out. Different factors affecting the optimization of the assay process were studied. Validation testes of the assay were carried out. The reproducibility as measured by the intra- and inter- assay variations is satisfactory. The recovery and dilution testes indicated accurate calibration and appropriate matrix. The present technique agreed well with the currently used commercial kit (Siemens, IRMA kit). In conclusion, the liquid phase double antibody RIA technique proved to be sensitive, specific, precis and accurate for routine laboratory use.

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Authors and Affiliations

  1. Labeled Compound Department, Hot Labs Center, Atomic Energy Authority, P. No. 13759, Cairo, Egypt

    H. M. Shafik, S. M. Ayoub, N. H. Ebeid & H. H. Someda

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  1. H. M. Shafik
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  2. S. M. Ayoub
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  3. N. H. Ebeid
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  4. H. H. Someda
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Correspondence to N. H. Ebeid.

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Shafik, H.M., Ayoub, S.M., Ebeid, N.H. et al. New adjuvant design using layered double hydroxide for production of polyclonal antibodies in radioimmunoassay techniques. J Radioanal Nucl Chem 301, 81–89 (2014). https://doi.org/10.1007/s10967-014-3155-5

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  • DOI: https://doi.org/10.1007/s10967-014-3155-5

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