Enzyme-linked immunosorbent assays using immune complexes for the diagnosis of tuberculosis

Arias-Bouda, Lenka M. Pereira; Kuijper, Sjoukje; Deutekom, H. van; Gijlswijk, Rob van; Pekel, Inge; Jansen, Henk M.; Kolk, A. H. J.

doi:10.1016/j.jim.2003.08.019

Cited by 18 publications

(5 citation statements)

References 15 publications

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“…7,8 Mycobacterial infection leads to the production of anti-LAM antibodies, the detection of which has been proposed for disease diagnosis. [17][18][19] Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

Structural Insights into Antibody Recognition of Mycobacterial Polysaccharides

Murase

Zheng

Joe

et al. 2009

Journal of Molecular Biology

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“…7,8 Mycobacterial infection leads to the production of anti-LAM antibodies, the detection of which has been proposed for disease diagnosis. [17][18][19] Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

Structural Insights into Antibody Recognition of Mycobacterial Polysaccharides

Murase

Zheng

Joe

et al. 2009

Journal of Molecular Biology

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“…Recombinant immunodominant proteins of infectious organisms are routinely employed in ELISA for the detection of disease-specific antibodies in the body fluids of suspected patients [ 5 , 6 , 17 , 31 – 33 ]. However, immobilization of these proteins on the surface of ELISA plates can be variable and inefficient [ 11 ].…”

Section: Resultsmentioning

confidence: 99%

Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection

et al. 2018

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The high-affinity interaction between biotin and streptavidin has opened avenues for using recombinant proteins with site-specific biotinylation to achieve efficient and directional immobilization. The site-specific biotinylation of proteins carrying a 15 amino acid long Biotin Acceptor Peptide tag (BAP; also known as AviTag) is effected on a specific lysine either by co-expressing the E. coli BirA enzyme in vivo or by using purified recombinant E. coli BirA enzyme in the presence of ATP and biotin in vitro. In this paper, we have designed a T7 promoter-lac operator-based expression vector for rapid and efficient cloning, and high-level cytosolic expression of proteins carrying a C-terminal BAP tag in E. coli with TEV protease cleavable N-terminal deca-histidine tag, useful for initial purification. Furthermore, a robust three-step purification pipeline integrated with well-optimized protocols for TEV protease-based H10 tag removal, and recombinant BirA enzyme-based site-specific in vitro biotinylation is described to obtain highly pure biotinylated proteins. Most importantly, the paper demonstrates superior sensitivities in indirect ELISA with directional and efficient immobilization of biotin-tagged proteins on streptavidin-coated surfaces in comparison to passive immobilization. The use of biotin-tagged proteins through specific immobilization also allows more efficient selection of binders from a phage-displayed naïve antibody library. In addition, for both these applications, specific immobilization requires much less amount of protein as compared to passive immobilization and can be easily multiplexed. The simplified strategy described here for the production of highly pure biotin-tagged proteins will find use in numerous applications, including those, which may require immobilization of multiple proteins simultaneously on a solid surface.

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“…Surprisingly, some nanobodies of the i ‐series showed specific binding to TAFIa, in spite of their selection towards intact TAFI. This apparent discrepancy could be explained by the possibility that TAFI, passively adsorbed to the microtiterplate, might undergo a conformational change (as observed for other antigens [29]) thereby displacing the activation peptide aside and making the catalytic cleft more accessible for nanobody interaction.…”

Section: Discussionmentioning

confidence: 99%

TAFIa inhibiting nanobodies as profibrinolytic tools and discovery of a new TAFIa conformation

Hendrickx

Winter

Buelens

et al. 2011

Journal of Thrombosis and Haemostasis

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Summary. Background: Because activated thrombin activatable fibrinolysis inhibitor (TAFIa) has very powerful antifibrinolytic properties, co-administration of t-PA and a TAFIa inhibitor enhances t-PA treatment. Objective: We aimed to generate nanobodies specifically inhibiting the TAFIa activity and to test their effect on t-PA induced clot lysis. Results: Five nanobodies, raised towards an activated more stable TAFIa mutant (TAFIa A ), are described. These nanobodies inhibit specifically TAFIa activity, resulting in an inhibition of up to 99% at a 16-fold molar excess of nanobody over TAFIa, IC 50 Õs range between 0.38-and > 16-fold molar excess. In vitro clot lysis experiments in the absence of thrombomodulin (TM) demonstrate that the nanobodies exhibit profibrinolytic effects. However, in the presence of TM, one nanobody exhibits an antifibrinolytic effect whereas the other nanobodies show a slight antifibrinolytic effect at low concentrations and a pronounced profibrinolytic effect at higher concentrations. This biphasic pattern was highly dependent on TM and t-PA concentration. The nanobodies were found to bind in the active-site region of TAFIa and their timedependent differential binding behavior during TAFIa inactivation revealed the occurrence of a yet unknown intermediate conformational transition. Conclusion: These nanobodies are very potent TAFIa inhibitors and constitute useful tools to accelerate fibrinolysis. Our data also demonstrate that the profibrinolytic effect of TAFIa inhibition may be reversed by the presence of TM. The identification of a new conformational transition provides new insights into the conformational inactivation of the unstable TAFIa.

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Enzyme-linked immunosorbent assays using immune complexes for the diagnosis of tuberculosis

Cited by 18 publications

References 15 publications

Structural Insights into Antibody Recognition of Mycobacterial Polysaccharides

Structural Insights into Antibody Recognition of Mycobacterial Polysaccharides

Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection

TAFIa inhibiting nanobodies as profibrinolytic tools and discovery of a new TAFIa conformation

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