Atypical antibody specificity: advancing the development of a generic assay for sulphonamides using heterologous ELISA

Abstract

Development of immunoassays with broad speci®city recognition of the sulphonamide drugs has proved surprisingly dif®cult in previous work. We have taken an antiserum speci®c for sulphachlorpyridazine in an ELISA system homologous for immunogen and solid phase conjugate and used it in a heterologous ELISA format. The assay proved to have broad speci®city, recognising 15 out of 21 drugs tested. Despite low cross-reactivity (<1%) for seven of the drugs tested, a fully optimised assay could prove useful as a multiresidue method for the group of drugs. The results provide evidence regarding the binding characteristics of the antibody and the immunogenicity of the target.

Disulfide Symmetric Dimers as Stable Pre‐Hapten Forms for Bioconjugation: A Strategy to Prepare Immunoreagents for the Detection of Sulfophenyl Carboxylate Residues in Environmental Samples

“…The possibility of tailor-made bioreceptors has been a matter of investigation mainly by using theoretical chemistry tools or molecular bioengineering methods. Thus, immunochemical methods have often been exploited for their capability to direct antibody specificity through appropriate hapten design by using theoretical methods and models [20,[36][37][38][39] or recombinant antibody methodologies. [40,41] However, as reported previously, addressing antibody recognition towards the common sulfonic group of the SPCs failed to produce generic antibodies.…”

“…[12,[15][16][17] The preparation of immunoreagents able to detect a wide range of short-alkyl-chain SPCs was one of the goals of this investigation. However, the production of antibodies with class-directed specificity is an uncertain goal that has been persecuted by many research groups with the purpose of developing immunochemical screening procedures able to detect, to a similar extent, different congeners of structurally related families of substances (i.e., sulfonamide antibiotics, [18][19][20] sulfonyl urea herbicides, [21,22] and so forth). The strategies employed frequently consist of preparing haptenprotein immunoconjugates that expose common features of the chemical structures to the immune system.…”

Application of computer-assisted molecular modeling for immunoassay of low molecular weight food contaminants: A review

“…CAMM not only can be useful in an iterative process producing feed-back information that can help improve hapten design, but it can also be useful in altering the ELISA assay format to improve assay performance. Studies that employed heterologous ELISA formats for development of specific assays for ceftiofur [31], nicarbazin [13], 2,4,6-trichlorophenol [93], and a group-specific assay for sulfonamides [94] were highly dependent on information gleaned from CAMM studies carried out on the targets of concern. It was determined that when heterologous ELISA formats were used in combination with CAMM studies, it was possible to improve assay performance in a more rapid manner.…”

Generic anti-sulfonamide immunoaffinity columns made using sulfamethazine-specific polyclonal antibodies

“…They are widely and routinely used in medicine and agriculture, and can persist as residues in food and the environment (Heberer 2002, Kolpin et al 2002. Several analytical techniques, such as enzyme-linked immunosorbent assay (ELISA) (Haasnoot et al 1996, Muldoon et al 2000a, Thomson & Sporns 1995, Haasnoot et al 2000, Spinks et al 2002, Lee et al 2001, Sheth & Sporns 1991 and immunoaffinity chromatography (Crabbe et al 1999, Li et al 2000, Grant et al 2003 rely upon antibodies to recognize sulfonamides. Although immunological techniques usually possess exquisite specificity, cross-reactivity is an inherent property of any antibody molecule toward antigens related to the immunogen.…”

“…Although immunological techniques usually possess exquisite specificity, cross-reactivity is an inherent property of any antibody molecule toward antigens related to the immunogen. Cross-reactivity is a positive characteristic for class specific or generic applications (Muldoon et al 2000b, Cliquet et al 2003, Assil et al 1992, Korpimaki et al 2004, Spinks et al 2002, Sheth & Sporns 1991, or is a negative characteristic in applications where specificity to a single compound within a class is desired (Lee et al 2001).…”