Lipid particles comprising variant paramyxovirus attachment glycoproteins and uses thereof
AssigneeSana Biotechnology, Inc.
InventorsPatricia CRUITE, Katherine Elizabeth SULLIVAN, Alexander Mark JUSTEN, Valeria BERLFEIN, Shirisha AMATYA, Albert Ruzo MATIAS, Kyle TRUDEAU, Matteo STOPPATO
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Abstract
Provided herein are lipid particles, including viral-based particles, comprising Paramyxovirus glycoproteins having reduced glycosylation, and in some aspects, comprise variant G/H proteins. Also provided are methods for producing and modifying lipid particles, including viral-based particles, wherein the particle has reduced glycosylation of the G/H protein, as well as methods for using the particles.
Claims87
A pseudotyped viral particle or virus-like particle comprising a Paramyxovirus glycoprotein (F) and a Paramyxovirus G/H glycoprotein, wherein the G/H protein has reduced glycosylation as compared to a wild-type G/H protein.
A lipid particle, comprising a lipid bilayer, a Paramyxovirus glycoprotein F, and a Paramyxovirus G/H glycoprotein, wherein the G/H protein is exposed on the outside of the lipid bilayer, and wherein the G/H protein has reduced glycosylation as compared to a wild-type G/H protein.
A variant paramyxovirus G/H protein comprising one or more amino acid substitutions at positions 39, 126, 128, 273, 345, 384, 448, and 496 corresponding to SEQ ID NO:
- 1
A polynucleotide comprising a nucleic acid molecule encoding a variant paramyxovirus G protein comprising an amino acid substitution at one or more amino acid positions that reduce glycosylation of the variant G/H protein.
A method of making a pseudotyped viral particle or virus-like particle comprising a paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising separating, enriching, or purifying the pseudotyped viral particle or virus-like particle, wherein the separating, enriching, or purifying further comprises enzymatic removal of glycans attached to the paramyxovirus F protein and/or paramyxovirus G/H protein.
A method of making a lipid particle comprising a paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising separating, enriching, or purifying the lipid particle, wherein the separating, enriching, or purifying further comprises enzymatic removal of glycans attached to the paramyxovirus F protein and/or paramyxovirus G/H protein.
A method of making a pseudotyped viral particle or virus-like particle comprising a paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising:
- a) providing a producer cell that comprises a viral particle or virus-like viral nucleic acid(s) and paramyxovirus F and/or paramyxovirus G/H nucleic acid(s);
- b) culturing the cell under conditions that allow for production of the viral particle or virus-like particle, wherein the culturing comprises treating the cell with a modulator of glycosylation;
- and
- c) separating, enriching, or purifying the viral particle or virus-like particle from the cell, thereby making the pseudotyped viral particle or virus-like particle.
A method of making a lipid particle comprising a paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising:
- a) providing a cell that comprises a paramyxovirus F and/or paramyxovirus G/H nucleic acid(s);
- b) culturing the cell under conditions that allow for production of a lipid particle, wherein the culturing comprises treating the cell with a modulator of glycosylation;
- and
- c) separating, enriching, or purifying the lipid particle from the cell, thereby making the lipid particle.
A composition comprising a producer cell, cell medium, and one or more modulators of glycosylation.
A method of making a pseudotyped viral particle or virus-like particle comprising a paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising:
- a) providing a producer cell that comprises a viral particle or virus-like viral nucleic acid(s) and paramyxovirus F and/or paramyxovirus G/H nucleic acid(s), wherein the producer cell comprises one or more genetic modifications that modulate glycosylation;
- b) culturing the cell under conditions that allow for production of the viral particle or virus-like particle, and
- c) separating, enriching, or purifying the viral particle or virus-like particle from the cell, thereby making the pseudotyped viral particle or virus-like particle.
A composition comprising a producer cell, wherein the producer cell comprises one or more nucleic acids for producing a viral particle or virus-like particle comprising a paramyxovirus F and/or paramyxovirus G/H protein, and wherein the producer cell comprises one or more genetic modifications that modulate glycosylation.
A method of making a lipid particle comprising a variant paramyxovirus F and/or paramyxovirus G/H protein having reduced glycosylation, comprising:
- a) providing a cell paramyxovirus F and/or paramyxovirus G/H nucleic acid(s), wherein the cell comprises one or more genetic modifications that modulate glycosylation;
- b) culturing the cell under conditions that allow for production of a lipid particle, and
- c) separating, enriching, or purifying the lipid particle from the cell, thereby making the lipid particle.
A composition comprising a producer cell, wherein the producer cell comprises one or more nucleic acids for producing a lipid particle comprising a paramyxovirus F and/or paramyxovirus G/H protein, and wherein the producer cell comprises one or more genetic modifications that modulate glycosylation.
Dependencies are inferred from the claim text.