V1/V2 fragments of a HIV-1 envelope glycoprotein

Patent metadata

Field	Value
Publication number	US-10507239-B2
Application number	US-201715635064-A
Country	US
Kind code	B2
Filing date	Jun 27, 2017
Priority date	Sep 11, 2012
Publication date	Dec 17, 2019
Grant date	Dec 17, 2019

Abstract

HIV-1 envelope proteins and fragments that possess naturally occurring and novel engineered epitopes that can be used to elicit (and are recognized by) broadly neutralizing antibodies.

First claim

The invention claimed is:

1. A method for producing a V1/V2 fragment of a HIV-1 envelope glycoprotein gp120 for use in eliciting broadly neutralizing antibodies, wherein the V1/V2 fragment binds to the broadly neutralizing PG9 monoclonal antibody (MAb), the method comprising:
expressing the V1/V2 fragment in a cell line lacking an N-acetylglucosaminyltransferase I (GnTI) enzyme activity, wherein the V1/V2 fragment comprises mannose-5 glycans and comprises the amino acid sequence as set forth in SEQ ID NO: 16.

2. The method of claim 1 , further comprising expressing a HIV-1 envelope glycoprotein, gp120, in a cell line lacking an N-acetylglucosaminyltransferase I (GnTI) enzyme activity and producing a formulation comprising the gp120 and the V1/V2 fragment, wherein mannose-5 glycan is incorporated in the gp120 and the V1/V2 fragment.

3. The method of claim 1 , wherein the V1/V2 fragment is expressed as a fusion protein comprising a signal sequence, a tag, and a linker.

4. The method of claim 2 , wherein the gp120 is a MN-rgp120 polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 wherein said MN-rgp120, when expressed in normal 293 cells, binds to PG9 and PGT128 MAb.

5. The method of claim 4 , wherein the MN-rgp120 comprises the amino acid sequence set forth in SEQ ID NO: 3.

6. The method of claim 1 , wherein the V1/V2 fragment is expressed as a fusion protein comprising a signal sequence, the 27 N-terminal amino acids of herpes simplex virus glycoprotein D, and a 3 amino acid linker (LLE).

7. The method of claim 2 , wherein the gp120 is A244-rgp120.

8. A formulation comprising a V1/V2 fragment of a HIV-1 envelope glycoprotein gp120 wherein the V1/V2 fragment comprises mannose-5 glycans and comprises the amino acid sequence as set forth in SEQ ID NO: 16.

9. The formulation of claim 8 , wherein the V1/V2 fragment is conjugated to a signal sequence.

10. The formulation of claim 8 , further comprising a HIV-1 envelope glycoprotein, A244-gp120, wherein mannose-5 glycan is incorporated in the A244-gp120.

11. The formulation of claim 8 , further comprising a HIV-1 envelope gp120 glycoprotein, wherein the gp120 is MN-rgp120 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, wherein said MN-rgp120, when expressed in normal 293 cells, binds to PG9 and PGT128 MAb.

12. The formulation of claim 11 , wherein the MN-rgp120 comprises the amino acid sequence set forth in SEQ ID NO: 3.

Assignees

• Univ California

Inventors

• Berman Phillip
• Tatsuno Gwen
• Yu Bin
• Morales Javier
• Mesa Kathryn

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P31/18

  for HIV · CPC title

• C12N2740/16122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K39/12 Primary

  Viral antigens · CPC title

• A61K39/21 Primary

  Retroviridae, e.g. equine infectious anemia virus · CPC title