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Myelin Basic Protein and a Multiple Sclerosis-related MBP-peptide Bind to Oligonucleotides.

Rozenblum GT, Kaufman T, Vitullo AD - Mol Ther Nucleic Acids (2014)

Bottom Line: Using purified MBP, we demonstrated the binding activity of the aptamers and we also showed the affinity of MBP for oligonucleotides of specific length.Moreover, one selected aptamer competitively inhibited the binding of an MBP-specific antibody to MBP and the aptamer was found more sensitive than a commercial antibody.In addition, we showed the ability of the aptamer to detect myelin-rich regions in paraffin-embedded mouse brain tissue.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Investigaciones Biomédicas y Biotecnológicas, Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y Diagnóstico, CEBBAD-Universidad Maimónides, Buenos Aires, Argentina.

ABSTRACT
Aptamer ligands for myelin basic protein (MBP) were obtained using the systematic evolution of ligand by exponential enrichment (SELEX) method. Two clones were isolated from a pool of oligonucleotides and tested for MBP targeting. Using purified MBP, we demonstrated the binding activity of the aptamers and we also showed the affinity of MBP for oligonucleotides of specific length. Moreover, one selected aptamer competitively inhibited the binding of an MBP-specific antibody to MBP and the aptamer was found more sensitive than a commercial antibody. In addition, we showed the ability of the aptamer to detect myelin-rich regions in paraffin-embedded mouse brain tissue. Therefore, the MBP-binding activity of the selected oligonucleotide may prove useful as a tool for life science and medical research for myelin detection and might be a good lead for testing it in autoimmune diseases such as multiple sclerosis.

No MeSH data available.


Related in: MedlinePlus

Detection of a myelin basic protein (MBP) peptide with MBPcl3 aptamer. Two micrograms of pure MBP peptide were incubated in a microtiter plate with equal nmol of an unselected aptamer library (Lib35) or MBPcl3. Bound aptamers were detected with HRP-conjugated avidin and developed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate (dark gray). Wells without MBP peptide were used as controls (light gray). Experiments were performed in triplicate. One asterisk indicates a significant difference in comparison with MBPcl3 and two asterisks indicates a significant difference in comparison with Lib35 (P < 0.05, Student's t-test). Data are shown as mean ± SD.
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fig6: Detection of a myelin basic protein (MBP) peptide with MBPcl3 aptamer. Two micrograms of pure MBP peptide were incubated in a microtiter plate with equal nmol of an unselected aptamer library (Lib35) or MBPcl3. Bound aptamers were detected with HRP-conjugated avidin and developed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate (dark gray). Wells without MBP peptide were used as controls (light gray). Experiments were performed in triplicate. One asterisk indicates a significant difference in comparison with MBPcl3 and two asterisks indicates a significant difference in comparison with Lib35 (P < 0.05, Student's t-test). Data are shown as mean ± SD.

Mentions: The recognition by T-cell receptors of antigenic peptides exposed on the surface of antigen presenting cells is considered an important step in MS progression. Thus, an MBP peptide comprising residues 90–106 (MBP90106) was synthesized and the MBP90106-binding activity of MBPcl3 was tested in an ELISA-like assay. Interestingly, MBPcl3 was able to bind MBP90106 as well as the unselected aptamer pool (Figure 6).


Myelin Basic Protein and a Multiple Sclerosis-related MBP-peptide Bind to Oligonucleotides.

Rozenblum GT, Kaufman T, Vitullo AD - Mol Ther Nucleic Acids (2014)

Detection of a myelin basic protein (MBP) peptide with MBPcl3 aptamer. Two micrograms of pure MBP peptide were incubated in a microtiter plate with equal nmol of an unselected aptamer library (Lib35) or MBPcl3. Bound aptamers were detected with HRP-conjugated avidin and developed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate (dark gray). Wells without MBP peptide were used as controls (light gray). Experiments were performed in triplicate. One asterisk indicates a significant difference in comparison with MBPcl3 and two asterisks indicates a significant difference in comparison with Lib35 (P < 0.05, Student's t-test). Data are shown as mean ± SD.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222649&req=5

fig6: Detection of a myelin basic protein (MBP) peptide with MBPcl3 aptamer. Two micrograms of pure MBP peptide were incubated in a microtiter plate with equal nmol of an unselected aptamer library (Lib35) or MBPcl3. Bound aptamers were detected with HRP-conjugated avidin and developed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate (dark gray). Wells without MBP peptide were used as controls (light gray). Experiments were performed in triplicate. One asterisk indicates a significant difference in comparison with MBPcl3 and two asterisks indicates a significant difference in comparison with Lib35 (P < 0.05, Student's t-test). Data are shown as mean ± SD.
Mentions: The recognition by T-cell receptors of antigenic peptides exposed on the surface of antigen presenting cells is considered an important step in MS progression. Thus, an MBP peptide comprising residues 90–106 (MBP90106) was synthesized and the MBP90106-binding activity of MBPcl3 was tested in an ELISA-like assay. Interestingly, MBPcl3 was able to bind MBP90106 as well as the unselected aptamer pool (Figure 6).

Bottom Line: Using purified MBP, we demonstrated the binding activity of the aptamers and we also showed the affinity of MBP for oligonucleotides of specific length.Moreover, one selected aptamer competitively inhibited the binding of an MBP-specific antibody to MBP and the aptamer was found more sensitive than a commercial antibody.In addition, we showed the ability of the aptamer to detect myelin-rich regions in paraffin-embedded mouse brain tissue.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Investigaciones Biomédicas y Biotecnológicas, Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y Diagnóstico, CEBBAD-Universidad Maimónides, Buenos Aires, Argentina.

ABSTRACT
Aptamer ligands for myelin basic protein (MBP) were obtained using the systematic evolution of ligand by exponential enrichment (SELEX) method. Two clones were isolated from a pool of oligonucleotides and tested for MBP targeting. Using purified MBP, we demonstrated the binding activity of the aptamers and we also showed the affinity of MBP for oligonucleotides of specific length. Moreover, one selected aptamer competitively inhibited the binding of an MBP-specific antibody to MBP and the aptamer was found more sensitive than a commercial antibody. In addition, we showed the ability of the aptamer to detect myelin-rich regions in paraffin-embedded mouse brain tissue. Therefore, the MBP-binding activity of the selected oligonucleotide may prove useful as a tool for life science and medical research for myelin detection and might be a good lead for testing it in autoimmune diseases such as multiple sclerosis.

No MeSH data available.


Related in: MedlinePlus