Approach to produce HIV-1 GP140 envelope protein trimers

摘要

An approach of producing recombinant trimers that mimic native HIV-1 envelope trimers is developed. A recombinant protein forming the recombinant trimers encompasses a recombinant HIV-1 gp140 fused to a tag through a linker at C-terminus of the recombinant HIV-1 gp140. The linker is sufficiently long so that the tag is accessible for binding by a binding molecule bound on a solid matrix. After expressed in a cell, the recombinant protein is secreted into the culture medium and assembles into recombinant trimers therein. The recombinant trimers may be directly purified from the culture medium. Cleaved and uncleaved trimers from different clade viruses are produced.

主权项

1. A recombinant DNA comprising a nucleic acid sequence encoding a recombinant protein,
wherein the recombinant protein comprises a recombinant HIV-1 gp140 fused to a tag through a peptide linker at C-terminus of the recombinant HIV-1 gp140, wherein the peptide linker is sufficiently long so that the tag is accessible for binding by a binding molecule bound on a solid matrix, wherein the recombinant protein assembles into recombinant trimers that mimic native HIV-1 envelope trimers when the recombinant protein is expressed by cells carrying the nucleic acid sequence encoding the recombinant protein, and wherein the recombinant protein comprises SEQ ID NO: 10.