Lipoparticles comprising proteins, methods of making, and using the same
The present invention relates to lipoparticles. The invention also relates to producing lipoparticles. The invention further relates to lipoparticles comprising a viral structural protein. The invention further relates to a lipoparticle comprising a membrane protein, and the lipoparticle can be attached to a sensor surface. The invention further relates to methods of producing and using the lipoparticle to, inter alia, assess protein binding interactions.
1. A method of identifying a binding partner of a multiple membrane spanning protein of interest comprising:
a) contacting a test binding partner with a lipoparticle, the lipoparticle comprising
an external lipid bilayer;
an enveloped retroviral structural protein; and
the multiple membrane spanning protein of interest,
wherein said enveloped retroviral structural protein is an uncleaved gag protein,
wherein said gag protein does not comprise a heterologous tag that binds to the multiple membrane spanning protein,
provided that the only viral proteins in the lipoparticle are structural proteins
b) detecting the presence of binding of the test binding partner to said membrane protein,
wherein the presence of binding identifies the test binding partner as a binding partner to said membrane protein of interest.
2. The method of claim 1 , wherein the test binding partner is expressed in a phage display library.
3. The method of claim 1 , wherein the test binding partner is present in a hybridoma supernatant.
4. The method of claim 1 , wherein said test binding partner is an antibody, antibody fragment, peptide, polypeptide, or small organic molecule.
5. The method of claim 1 , wherein the test binding partner is a monoclonal antibody, a polyclonal antibody, an affinity-purified polyclonal antibody, a Fab fragment derived from a monoclonal antibody, an immunoglobulin-fusion protein, a single-chain Fv, or an Fc-fusion protein.
6. The method of claim 1 , wherein the multiple membrane spanning protein of interest is a G-protein coupled receptor.
7. The method of claim 1 , wherein the multiple membrane spanning protein of interest is an ion channel.
8. The method of claim 1 , wherein the multiple membrane spanning protein of interest is a transporter protein.
9. The method of claim 1 , wherein the multiple membrane spanning protein of interest is a glucose transporter protein.
10. The method of claim 1 , wherein the glucose transporter protein is Glut4.
11. The method of claim 1 , wherein the multiple membrane spanning protein of interest is an amino acid transporter.
12. The method of claim 1 , wherein the multiple membrane protein of interest is selected from the group consisting of: CCR4, CCR5, CCR7, CCR8, CX3CR1, APJ, XCR1 Glut4, Kv1.3, MCAT1, CFTR, Shaker, CXCR4, mu-opioid receptor, KCNH2, and Presinilin-1.
13. A method of identifying a binding partner of a multiple membrane spanning protein of interest comprising:
a) contacting a library of test binding partners with a lipoparticle, the lipoparticle comprising:
an external lipid bilayer;
an enveloped retroviral structural protein; and
the multiple membrane spanning protein of interest,
wherein said enveloped retroviral structural protein is an uncleaved gag protein,
wherein said gag protein does not comprise a heterologous tag that binds to the multiple membrane spanning protein,
provided that the only viral proteins in the lipoparticle are structural proteins
b) detecting the presence of binding of the test binding partner to said membrane protein,
wherein the presence of binding identifies a member of the library as a binding partner to said membrane protein of interest.
14. The method of claim 13 , wherein the library is a phage display library.
15. The method of claim 13 , wherein the library is a library of hybridoma supernatants.
16. The method of claim 13 , wherein said library of test binding partners is a library of antibodies, antibody fragments, peptides, polypeptides, or small organic molecules.
17. The method of claim 13 , wherein the library of test binding partners is a library of monoclonal antibodies, polyclonal antibodies, affinity-purified polyclonal antibodies, Fab fragments, immunoglobulin-fusion proteins, single-chain Fv, or Fc-fusion proteins.
18. The method of claim 13 , wherein the multiple membrane spanning protein of interest is a G-protein coupled receptor, an ion channel or a transporter protein.
19. The method of claim 13 , wherein the multiple membrane spanning protein of interest is a glucose transporter protein.
20. The method of claim 13 , wherein the glucose transporter protein is Glut4.