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Inventor(s): Richard James Reister (Plano, TX) Assignee(s): The Modern Gentleman, Inc. (Plano, TX) Law Firm: Slater Matsil, LLP (Local + 1 other metros) Application No., Date, Speed: 29610470 on 07/12/2017 (517 days app to issue)

[A61K] PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J 3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

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After partially blocking the Abs sites on the nanoprobes with PD-L1, the decrease in intensity (ΔI) versus the amount of PD-L1 (m) showed a linear relation in the range 1.0 to 5.0 fmol (R = 0.999, n = 5; Fig. 5G)

We further investigated the PD-L1 expression on TNBC surfaces using Raman mapping. We collected a topographic image of 50 μm by 50 μm from the integrated intensity of the peak at 1530 cm−1. The first column of Fig. 5L shows the Raman mapping images of the PD-L1 expression level of three types of TNBCs. The color-false images revealed that the expression level of PD-L1 on three types of TNBCs was MDA-MB-231 > HCC38 >> MCF-7. Both the calculated results and Raman mapping images were consistent with those of the commercial enzyme-linked immunosorbent assay (ELISA) kit (fig. S7B). The Raman imaging technique mentioned above was further successfully used to monitor dynamic PD-L1 expression on three different TNBCs in response to IFN-γ, which was a type of biomolecule reported to upgrade the expression of PD-L1 on cell surfaces (38). Three types of IFN-γ–treated TNBCs progressively increased with the concentrations of IFN-γ (Fig. 5, M to P). Besides concentration-dependent mapping, to provide additional evidence that our probe selectively targets PD-L1 on cell surfaces with high sensitivity, we further conducted time-dependent Raman mapping experiments for IFN stimulation. Figure S8 shows the time-dependent responses for IFN-γ during the course of the experiment. The Raman peak obtained from IFN-γ–treated cells increased as the IFN-γ incubation time increased and reached a plateau after 48 hours. Both the histogram and Raman mapping images indicated that IFN-γ treatment significantly stimulated the expression of PD-L1 on cell surfaces within 48 hours. All these results confirmed the high sensitivity and selectivity of the MGT-Abs-CuPc probe for Raman bioanalysis and imaging.

[G06F] ELECTRIC DIGITAL DATA PROCESSING (computer systems based on specific computational models G06N)

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