Gel Filtration Separation of ICG and NPs

To separate free ICG and NPs, a gel filtration procedure was developed by modifying literature methods [43 (link),44 (link)]. Here, 2 g Sephadex G-25 (Fine, Sigma Aldrich, Milan, Italy) was hydrated in distilled water, and a chromatographic column was packed by gravity. A flux of 0.4 mL/min was maintained by means of an HPLC pump (isocratic Kontron 420, Kontron Instruments, Milan, Italy). Then, 250 µL of ICG aqueous solution 0.4 M was loaded as reference. Subsequently, 1 mL fractions were collected and analyzed at 780 nm wavelength by means of a spectrophotometer (Perkin Elmer Lambda 25, Milan, Italy) versus a calibration curve. Different amounts of NPs were mixed with the same amount of ICG solution of the reference experiment and loaded in the column, obtaining molar ratios between ICG and CS (as a monomeric unit) ranging from 8:1 to 0.4:1. In these cases, as well, 1 mL volumes were collected and analyzed for ICG concentration. A similar procedure was followed when loading ads-ICG NPs and enc-ICG NPs, both at an ICG:CS molar ratio of 0.4. These were, however, eluted not only with distilled water but also with DMEM cell culture medium supplemented with 10% v/v inactivated fetal calf bovine serum (Euroclone, Milan, Italy).

Free full text: Click here

Miele D., Sorrenti M., Catenacci L., Minzioni P., Marrubini G., Amendola V., Maestri M., Giunchedi P, & Bonferoni M.C. (2022). Association of Indocyanine Green with Chitosan Oleate Coated PLGA Nanoparticles for Photodynamic Therapy. Pharmaceutics, 14(8), 1740.

Publication 2022

Sephadex G-25 (Fine, Lambda 25

Cell Cell culture Chromatographic Culture medium Fetal bovine serum Gel filtration Gravity Hplc Molar Sephadex

Corresponding Organization : University of Pavia

Other organizations : Policlinico San Matteo Fondazione, Istituti di Ricovero e Cura a Carattere Scientifico, University of Sassari

Top 5 similar protocols

Variable analysis

independent variables

Molar ratio between ICG and CS (as a monomeric unit) (ranging from 8:1 to 0.4:1)
Type of nanoparticles (ads-ICG NPs and enc-ICG NPs)

dependent variables

ICG concentration in collected fractions

control variables

Amount of ICG solution loaded as reference (250 µL of 0.4 M ICG aqueous solution)
Flux rate maintained at 0.4 mL/min using an HPLC pump
Wavelength used for spectrophotometric analysis (780 nm)
Chromatographic column (2 g Sephadex G-25 (Fine, Sigma Aldrich, Milan, Italy) packed by gravity)
Elution solvent (distilled water, and DMEM cell culture medium supplemented with 10% v/v inactivated fetal calf bovine serum)

controls

Positive control: 250 µL of 0.4 M ICG aqueous solution loaded as reference
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!