Advances in Immunoassay Technology by N. Chiu, et. al.

Advances in Immunoassay Technology by N. Chiu, et. al.

By N. Chiu, et. al.

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Surface biofunctionalization of materials by amine groups. Journal of Material Research, Vol. 19 No. 8, pp. , Takkinen, K. (2005). One step homogenous immunoassays for small analytes. Analytical Chemistry, Vol. 77, No. 7, pp. 2637-2642 Rejeb, S. , Khonsari, F. A. and Durand, N. F. (1998). Functionalization of nitrocellulose membranes using ammonia plasma for the covalent attachment of antibodies for use in membrane-based immunoassays. Analytica Chimica Acta, Vol. 376, No. 1, (4 December 1998), pp.

Heterogeneous assays such as Enzyme Linked Immuno-Sorbent Assays (ELISAs) are most widely used detection method which utilizes the concept of immobilization of biomolecules on solid support. These have atleast one separation step allowing the differentiation of reacted from un-reacted materials. The enzymatic activity is quantified either in bound state or free fraction by an enzyme catalyzed process of a relatively nonchromatic substrate to highly chromatic product. ,1976]. In ELISA, antibody immobilized on the solid support detects the specific antigen present in the sample and this immune complex is detected by a high turn-over enzyme conjugated antibody.

For these reasons there may be a higher prevalence of unpredictable crossreaction in IMAs than in the single-site antigen-antibody reaction in reagent-limited assays (Boscato & Stuart, 1986). , 1997). Exogenous interferences are any interference caused by the introduction of external factors or conditions, in vivo or in vitro, not normally present in native, properly collected and stored samples. For example, hemolysis, lipemia, icterus, blood collection tube additives, Fig. 1. Different interferences in immunometric immunoassays: Aa - assay without any interference; Ab - cross-reactivity of an interfering substance with capture antibody, resulting in false negative result; B - positive interference: Ba - unspecific binding of labelled detector antibody to a not blocked solid phase; Bb - “bridge” binding by heterophilic antibodies or HAMA, respectively; C - negative interference: Ca - change of sterical conformation after binding of interfering protein to Fc fragment of detector antibody Cb masking of the epitope on analyte surface by a protein of the sample (Dodig, 2009).

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