Production of soluble hiv envelope trimers in planta
AssigneeUniversity of Cape Town
InventorsAnn Elizabeth Meyers, Anna-Lise Williamson, Edward Peter Rybicki, Emmanuel Aubrey Margolin, Rosamund Chapman
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Abstract
The present invention relates to a method for producing a recombinant HIV glycoprotein polypeptide in a plant and to trimeric complexes of the recombinant, plant-produced HIV glycoprotein polypeptide which mimic the native HIV Env complex. The invention also relates to nucleic acids encoding the recombinant polypeptides, expression vectors containing the aforementioned nucleic acids and to pharmaceutical compositions, uses and methods of eliciting an immune response against HIV in a subject using the recombinant polypeptides and trimeric complexes.
Claims20
A method for producing a recombinant polypeptide capable of forming a trimeric Env glycoprotein complex, the method comprising the steps of:
- (i) providing a codon-optimised nucleotide sequence encoding a recombinant polypeptide having the formula:
- X1—X2—X3—X4
- wherein,
- X1 is a secretory signal peptide;
- X2 is an HIV gp120 envelope polypeptide;
- X3 is a linker peptide;
- and
- X4 is a HIV gp41 polypeptide,
- wherein the gp41 polypeptide is selected from either a full-length gp41 polypeptide or a truncated gp41 polypeptide, and wherein if the gp41 polypeptide is a truncated gp41 polypeptide then it is truncated after the LALD motif of the gp41 polypeptide;
- further wherein the recombinant polypeptide comprises an I559P mutation;
- (ii) cloning the codon-optimised nucleic acid encoding the recombinant polypeptide into an expression vector adapted to express the recombinant polypeptide in a plant cell;
- (iii) transforming the plant cell with the expression vector of step (ii);
- (iv) expressing the recombinant polypeptide in the plant cell;
- and
- (v) recovering the recombinant polypeptide from the plant cell.
Dependencies are inferred from the claim text.