Evaluation of the concept of heterology in a monoclonal antibody-based ELISA utilizing direct hapten linkage to polystyrene microtiter plates

Holthues, Heike; Pfeifer-Fukumura, Ursula; Sound, Iris; Baumann, Wolfram

doi:10.1016/j.jim.2005.06.012

Cited by 28 publications

(18 citation statements)

References 22 publications

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“…The reaction mixture was diluted with benzene and washed four times with cool water and once with brine, dried over anhydrous Na 2 SO 4 , and concentrated under vacuum to give the crude product that was purified by chromatography, using CHCl 3 as eluent, to give hapten PCb6 (2) (67 mg, 75%) as a viscous oil. 1 adjuvant. Whole blood was collected from the ear vein of the rabbits and by intracardiac puncture 10 days after the fourth injection.…”

Section: Synthesis Of (E)-6-(2-(2-(13-dimethoxy-3-oxoprop-1-en-2-yl)mentioning

confidence: 99%

Exploring alternative hapten tethering sites for high-affinity anti-picoxystrobin antibody generation

Parra¹,

Esteve‐Turrillas²,

Abad‐Somovilla³

et al. 2011

Analytical Biochemistry

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The relevance of the linker tethering site in haptens was investigated for antibody generation and immunoassay development. Three derivatives of the strobilurin fungicide picoxystrobin were synthesized with the same functionalized spacer arm located at three different positions. Protein conjugates of those haptens were employed as immunogens and novel polyclonal antibodies were produced and characterized. All haptens afforded highly specific antibodies, but different affinities to the free analyte were observed among the obtained antisera. Then, competitive enzyme-linked immunosorbent assays were studied in several formats, and site heterology was confirmed as an effective strategy for detectability improvement. An indirect heterologous immunoassay was eventually selected and optimized, showing a limit of detection for picoxystrobin of 0.02 µg/L and a working range between 0.03 and 1.30 µg/L. Finally, the developed extraction and analytical procedures revealed a practical limit of quantification of 5 µg/kg for this fungicide in soybean sprouts, well below the maximum residue limits in the EU.

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Section: Synthesis Of (E)-6-(2-(2-(13-dimethoxy-3-oxoprop-1-en-2-yl)mentioning

confidence: 99%

Exploring alternative hapten tethering sites for high-affinity anti-picoxystrobin antibody generation

Parra¹,

Esteve‐Turrillas²,

Abad‐Somovilla³

et al. 2011

Analytical Biochemistry

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“…In heterologous assays, the immunizing and competitor haptens differ in their structure, whereas, in the homologous assays, the haptens are the same; the assay sensitivity of such heterologous assays is reported to be better than in homologous assay set-ups [18,[46][47][48]. KLH-EDC-his was used as probe as well as to generate Ab1; thus, expectedly, the affinity of Ab1 was high towards the protein-EDC-his conjugates.…”

Section: Discussionmentioning

confidence: 99%

Challenges in Developing a Biochip for Intact Histamine Using Commercial Antibodies

2017

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This study describes the development and the challenges in the development of an on-chip immunoassay for histamine using commercially available antibodies. Histamine can be used as an indicator of food freshness and quality, but it is also a relevant marker in clinical diagnostics. Due to its low molecular weight, simple structure and thus low immunogenicity production of high specificity and affinity antibodies is difficult. From six commercial anti-histamine antibodies tested, only two bound the histamine free in the solution. A fluorescent on-chip immunoassay for histamine was established with a dynamic range of 8-111 µg/mL using polyclonal anti-histamine antibody H7403 from Sigma (Mendota Heights, MN, USA). The anti-histamine antibodies described and used in published literature are thoroughly reviewed and the quality of commercial antibodies and their traceability and quality issues are highlighted and extensively discussed.

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“…Moreover, different authors supported that direct hapten coated format showed an improvement in immunoassay sensitivity as well; this probably due to changes in the valency of antibody-hapten interaction (Gerdes et al, 1999;Townsend et al, 2006). In line with this, several approaches have been developed on modified polystyrene (PS) microtriter wells of ELISA plates, resulting in improved analytical performances of the assays (Böcher and Sorensen, 1994;Feng et al, 2009;Kaur et al, 2008;Holthues et al, 2005;Niveleau et al, 1993).…”

Section: Introductionmentioning

confidence: 89%

Direct hapten-linked multiplexed immunoassays on polycarbonate surface

Tamarit-López¹,

Morais²,

Puchades³

et al. 2011

Biosensors and Bioelectronics

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Direct hapten-linked multiplexed immunoassay is developed on the polycarbonate surface of standard Digital Versatile Discs (DVDs) for six compounds of environmental concern, as proof of concept. Carboxylated haptens are directly linked to the aminated polycarbonate surface through carbodiimide/succinimide coupling. The modified DVD surface maintained its physical and optical properties. Multiplexed assay reached detection limits down to 0.1 μg/L for chlorpyrifos, 2,4,5-trichlorophenoxypropionic acid, sulfathiazole and sulfasalazine and down to 1.0 μg/L for fenthion and malathion. This approach presents advantages such as the improvement in sensitivity in comparison to protein-hapten conjugate format for all the studied analytes and the absence of cross-interference effects, allowing high throughput multianalysis on the same surface. Also, a comparison of the performance of two sensing strategies indicated that DVD disc and drive approach turned out in a simpler mode, the assays being more reproducible and with higher signal to noise ratios.

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Evaluation of the concept of heterology in a monoclonal antibody-based ELISA utilizing direct hapten linkage to polystyrene microtiter plates

Cited by 28 publications

References 22 publications

Exploring alternative hapten tethering sites for high-affinity anti-picoxystrobin antibody generation

Exploring alternative hapten tethering sites for high-affinity anti-picoxystrobin antibody generation

Challenges in Developing a Biochip for Intact Histamine Using Commercial Antibodies

Direct hapten-linked multiplexed immunoassays on polycarbonate surface

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