Journal: Nanoscale

Article Title: Site-Specific Conjugation of Antibody on Gold Nanoparticle Surface for One-Step Diagnosis of Prostate Specific Antigen with Dynamic Light Scattering

doi: 10.1039/c7nr03096g

Figure Lengend Snippet: Absorbance spectra of AuNPs before (black line) and after functionalization with two different PSA specific antibodies: A) B731M and B) B728M via the ionic interaction, thiol reduction, EDC-NHS and UV-NBS methods, indicating successful functionalization of AuNPs with antibodies for all four methods without causing any aggregation. C) DLS measurements of AuNPs (purple bar), non-functionalized antibodies, and PSA specific antibody functionalized AuNPs via the UV-NBS, EDC-NHS, thiol reduction and ionic interaction methods.

Article Snippet: Purified free prostate specific antigen (PSA), mouse anti-PSA (clone: B731M), and mouse anti-PSA (clone: B728M) were purchased from Meridian Life Science, Inc. (Memphis, TN).

Techniques:

Journal: Nanoscale

Article Title: Site-Specific Conjugation of Antibody on Gold Nanoparticle Surface for One-Step Diagnosis of Prostate Specific Antigen with Dynamic Light Scattering

doi: 10.1039/c7nr03096g

Figure Lengend Snippet: A) Schematic representation of antibody functionalized AuNPs’ cluster formation upon binding of antigen. Antigen detection sensitivity of the antibody functionalized AuNPs prepared utilizing the B) ionic interaction, C) thiol reduction, D) EDC-NHS, and E) UV-NBS methods, was determined with DLS measurements. Size of the antibody functionalized AuNPs was measured with DLS upon addition of increasing concentrations of PSA. As expected, the size of AuNPs functionalized with only one antibody, either B731M (blue bar) or B728M (red bar), did not change whereas mixture of B731M and B728M functionalized AuNPs (green bar) formed clusters upon addition of increasing concentration of PSA (F) Comparison of cluster formations (green bars from B, C, D and E) utilized with all methods: UV-NBS (purple), EDC-NHS (blue), Thiol Reduction (orange), and Ionic Interaction (green). All data represent means (±SD) of triplicate experiments.

Article Snippet: Purified free prostate specific antigen (PSA), mouse anti-PSA (clone: B731M), and mouse anti-PSA (clone: B728M) were purchased from Meridian Life Science, Inc. (Memphis, TN).

Techniques: Binding Assay, Concentration Assay, Comparison

Journal: Nanoscale

Article Title: Site-Specific Conjugation of Antibody on Gold Nanoparticle Surface for One-Step Diagnosis of Prostate Specific Antigen with Dynamic Light Scattering

doi: 10.1039/c7nr03096g

Figure Lengend Snippet: The specificity of the AuNP detection system prepared via the UV-NBS method was tested by measuring the cluster formation of AuNPs that are functionalized with PSA specific antibodies; B731M and B728M in the presence of non-specific targets such as A) BSA, B) transferrin, and C) hemoglobin instead of PSA. As expected, cluster formation was not observed, with non-specific protein targets utilizing anti-PSA immobilized AuNPs.

Article Snippet: Purified free prostate specific antigen (PSA), mouse anti-PSA (clone: B731M), and mouse anti-PSA (clone: B728M) were purchased from Meridian Life Science, Inc. (Memphis, TN).

Techniques:

Journal: Nanoscale

Article Title: Site-Specific Conjugation of Antibody on Gold Nanoparticle Surface for One-Step Diagnosis of Prostate Specific Antigen with Dynamic Light Scattering

doi: 10.1039/c7nr03096g

Figure Lengend Snippet: AuNPs were conjugated with Rituximab, anti-CD20 antibody, to validate the selectivity of the antibody functionalized AuNP detection system against to PSA target. Sandwich assays were carried out utilizing one of anti-PSA antibody functionalized AuNPs, either A) B731M or B) B728M, and Rituximab functionalized AuNP to target PSA. Results show that there are no non-specific interactions.

Article Snippet: Purified free prostate specific antigen (PSA), mouse anti-PSA (clone: B731M), and mouse anti-PSA (clone: B728M) were purchased from Meridian Life Science, Inc. (Memphis, TN).

Techniques: