Identification and elimination of false-positives in an ELISA-based system for qualitative assessment of

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Babak Afrough1, Miriam V Dwek, Pamela Greenwell

  • 1University of Westminster, London, UK.

Biotechniques
|November 21, 2007
PubMed

Systematic optimization of a lectin-based enzyme-linked immunosorbent assay (ELISA) procedure using a panel of 21 biotinylated plant lectins and a glycoprotein with defined glycosylation (i) identified blocking as a limiting step in solid phase sugar binding analysis and (ii) found Synblock to be a better alternative to bovine serum albumin (BSA) as blocking agent.

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Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.