Methods for profiling extracellular vesicles from combinatorial phage display libraries
A method for profiling extracellular vesicles (EV) is provided. The method includes: (a) incubating isolated EV from one or more sources with a phage display library under conditions suitable for forming EV-bound phage, wherein the phage display library comprises one or more types of phage, each type of phage having a displayed proteinaceous display binding moieties capable of recognizing one or more epitopes on the surface of the EV, wherein each type of phage comprises nucleic acids encoding the displayed proteinaceous display binding moieties, and wherein the displayed proteinaceous display binding moieties of each type of phage is different; (b) isolating the EV-bound phage; (c) extracting the nucleic acids from the EV-bound phage; (d) amplifying the extracted nucleic acids; (e) sequencing the amplified nucleic acids to identify specific nucleic acid sequences associated with each type of phage from the library that are enriched from the isolated EV; and (f) comparing the specific nucleic acid sequences (output) with nucleic acid sequences from the phage display library (input) and nucleic acid sequences from a control to identify sequences that can distinguish between EV from the one or more sources.
1. A method for profiling extracellular vesicles (EVs), the method comprising:
incubating isolated EVs from one or more sources with a phage display library under conditions suitable for forming EV-bound phage complexes, wherein the phage display library comprises one or more types of phage, each type of phage having a displayed Dab, Fab, or scFv capable of binding one or more epitopes on the surface of the EVs, wherein each type of phage comprises nucleic acids encoding the displayed Dab, Fab, or scFv, and wherein the displayed Dab, Fab, or scFv of each type of phage is different;
isolating the EV-bound phage complexes;
extracting the nucleic acids from the EV-bound phages;
amplifying the extracted nucleic acids;
sequencing the amplified nucleic acids to identify specific nucleic acid sequences from the EV-bound phages; and
identifying the epitopes present on the surface of the EVs of the EV-bound phage complexes.
2. The method according to claim 1 , wherein the isolated EVs are obtained from a bodily fluid.
3. The method according to claim 1 , wherein the isolated EVs are obtained from cell lines, serum, plasma, urine, saliva, or cerebral fluid.
4. The method according to claim 1 , wherein the EV-bound phage complexes are isolated by immunoprecipitation.
5. The method according to claim 1 , wherein the EV-bound phage complexes are isolated by polymer precipitation.
6. The method according to claim 1 , wherein the EVs are isolated by immunoprecipitation, ultracentrifugation, or affinity purification.
7. The method according to claim 1 , wherein M13 phage is used to generate the phage display library.