DNP-BSA is a multivalent antigen and protein-binding reagent. DNP-BSA binds to subdomain IIA (Site I) of bovine serum albumin via predominantly hydrophobic forces, with spontaneous complex formation via combined dynamic and static quenching. DNP-BSA induces cross-linking of IgE-bound FceRI, triggering tyrosine phosphorylation of FceRI β and γ subunits. DNP-BSA binds to anti-DNP IgE via its DNP groups, forming cross-links between IgE-FceRI complexes, with most DNP groups initially unavailable but transiently exposed for binding. DNP-BSA induces FITC fluorescence quenching in FITC-labeled anti-DNP IgE, proportional to occupied IgE Fab binding sites. DNP-BSA can be used for the research of allergy, inflammation, and contact sensitivity (allergic contact dermatitis ) [1] [2] [3].
In Vivo, DNP-BSA (3-100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) potently suppresses DNFB-induced contact sensitivity in BALB/c mice, reducing key inflammatory parameters by 54-81% and inducing TCRab+ CD4+ CD25+ FoxP3+ regulatory T cells that mediate suppression[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) suppresses DNFB-induced contact sensitivity and induces TCRab+ CD4+ CD25+ FoxP3+ regulatory T cells in C57BL/6 mice[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) induces regulatory lymph node cells that potently suppress DNFB-induced contact sensitivity in B10.PL and related knockout mice, with suppression efficacy ranging from 80-92%[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) induces regulatory T cells that distribute equally across peripheral lymphoid tissues and suppress DNFB-induced contact sensitivity in NOD mice[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) suppresses DNFB-induced lymph node cell proliferation by 43-56% and reduces proinflammatory cytokine production in BALB/c mice[3].
In Vitro, DNP-BSA forms spontaneously under physiological pH, with hydrophobic forces as the dominant driving force, as evidenced by positive ΔH° (46.22 kJ mol-1) and ΔS° (237.2 J mol-1 K-1) values and negative ΔG° values ranging from -24.46 to -29.21 kJ mol-1[1]. DNP-BSA forms at site I of BSA's subdomain IIA, forming stable interactions with Trp213, Leu197, Arg217, Asp450, and Leu480, with a docking score of -8.71 kcal/mol[1]. Stimulates FceRI β subunit tyrosine phosphorylation in RBL-2H3 cells that correlates directly with the kinetics of DNP-BSA-mediated IgE-FceRI cross-linking at 15°C, but shows a biphasic decline while cross-linking increases at 35°C[2]. DNP-BSA potently stimulates degranulation of adherent RBL-2H3 cells, inducing ~50% β-hexosaminidase release at concentrations of 0.2 nM and 1.1 nM[2].
References:
[1]. El Gammal RN, et al. Insights on the in-vitro binding interaction between donepezil and bovine serum albumin. BMC chemistry. 2023 Apr 06;17(1):31.
[2]. Xu K, et al. Kinetics of multivalent antigen DNP-BSA binding to IgE-Fc epsilon RI in relationship to the stimulated tyrosine phosphorylation of Fc epsilon RI. J Immunol. 1998 Apr 1;160(7):3225-35.
[3]. Majewska-Szczepanik M, et al. Epicutaneous immunization with DNP-BSA induces CD4+ CD25+ Treg cells that inhibit Tc1-mediated CS. Immunology and cell biology. 2012 Sep;90(8):784-95.
In Vivo, DNP-BSA (3-100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) potently suppresses DNFB-induced contact sensitivity in BALB/c mice, reducing key inflammatory parameters by 54-81% and inducing TCRab+ CD4+ CD25+ FoxP3+ regulatory T cells that mediate suppression[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) suppresses DNFB-induced contact sensitivity and induces TCRab+ CD4+ CD25+ FoxP3+ regulatory T cells in C57BL/6 mice[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) induces regulatory lymph node cells that potently suppress DNFB-induced contact sensitivity in B10.PL and related knockout mice, with suppression efficacy ranging from 80-92%[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) induces regulatory T cells that distribute equally across peripheral lymphoid tissues and suppress DNFB-induced contact sensitivity in NOD mice[3]. DNP-BSA (100 µg per animal; epicutaneous, via occlusive patch; days 0 and 4, each patch left in place for 3 days) suppresses DNFB-induced lymph node cell proliferation by 43-56% and reduces proinflammatory cytokine production in BALB/c mice[3].
In Vitro, DNP-BSA forms spontaneously under physiological pH, with hydrophobic forces as the dominant driving force, as evidenced by positive ΔH° (46.22 kJ mol-1) and ΔS° (237.2 J mol-1 K-1) values and negative ΔG° values ranging from -24.46 to -29.21 kJ mol-1[1]. DNP-BSA forms at site I of BSA's subdomain IIA, forming stable interactions with Trp213, Leu197, Arg217, Asp450, and Leu480, with a docking score of -8.71 kcal/mol[1]. Stimulates FceRI β subunit tyrosine phosphorylation in RBL-2H3 cells that correlates directly with the kinetics of DNP-BSA-mediated IgE-FceRI cross-linking at 15°C, but shows a biphasic decline while cross-linking increases at 35°C[2]. DNP-BSA potently stimulates degranulation of adherent RBL-2H3 cells, inducing ~50% β-hexosaminidase release at concentrations of 0.2 nM and 1.1 nM[2].
References:
[1]. El Gammal RN, et al. Insights on the in-vitro binding interaction between donepezil and bovine serum albumin. BMC chemistry. 2023 Apr 06;17(1):31.
[2]. Xu K, et al. Kinetics of multivalent antigen DNP-BSA binding to IgE-Fc epsilon RI in relationship to the stimulated tyrosine phosphorylation of Fc epsilon RI. J Immunol. 1998 Apr 1;160(7):3225-35.
[3]. Majewska-Szczepanik M, et al. Epicutaneous immunization with DNP-BSA induces CD4+ CD25+ Treg cells that inhibit Tc1-mediated CS. Immunology and cell biology. 2012 Sep;90(8):784-95.