Back to results
US 11,298,415 B2Granted
Alginate chitosan nanoformulation of OmpA—a Shigella protein subunit
AssigneeIndian Council of Medical Research
InventorsHemanta Koley, Priyadarshini Mukherjee, Dhrubajyoti Nag, Ritam Sinha, Manoj Kumar Chakrabarti
Life of this patent
Estimating the term…
Abstract
Provided herein is a Formulation of OmpA including a) OmpA protein as active molecule obtained as a product of OmpA gene inserted in a plasmid comprising a novel set of forward and reverse primer set, and b) Alginate chitosan nanoparticles as vehicle.
Claims7
An oral formulation of outer membrane protein A (OmpA) comprising essentially of:
- a) OmpA protein as an active molecule, obtained as a product of an OmpA gene derived from Shigella flexneri 2a 2457T, inserted in a plasmid comprising a set of a forward and a reverse primer set;
- and
- b) Alginate chitosan nanoparticles as a vehicle;
- wherein the formulation has a particle diameter in a range of 240 nm to 700 nm;
- wherein the formula is administered orally;
- wherein the formulation has a mean particle size of 535.0 nm, a particle size distribution of 0.470 and a zeta potential of −20.78;
- wherein the forward primer has a nucleotide sequence 5′-AAAAAGACACATATG CG ATT GCAG-3′ (SEQ ID NO:
- 1) and the reverse primer has a nucleotide sequence 5′-GTATCTCGAGAGCTTGCGCTGAGTTAC-3′ (SEQ ID NO:
- 2);
- and,
- wherein bases “CATATG” in the forward primer represents restriction site specific for NcoI and the bases “CTCGAG” in the reverse primer represents restriction site specific for XhoI.
8 more elements
A method of preparation of an oral formulation of an outer membrane protein A (OmpA), the method comprising the steps:
- a) preparing a recombinant plasmid comprising an OmpA forward primer having a nucleotide sequence 5′-AAAAAGACACATATG CG ATT GCAG-3′ (SEQ ID NO:
- 1) and an OmpA reverse primer having a nucleotide sequence
- 5′-GTATCTCGAGAGCTTGCGCTGAGTTAC-3′ (SEQ ID NO:
- 2)
- wherein the bases “CATATG” in the forward primer represents restriction site specific for NcoI and the bases “CTCGAG” in the reverse primer represents restriction site specific for XhoI;
- b) transferring the recombinant plasmid to E. coli BL21 (DE3) for multiplication;
- c) isolating OmpA protein as a product from the E. coli BL21 (DE3);
- d) preparing chitosan with alginate nanoparticles;
- and
- e) loading OmpA protein into said chitosan with alginate nanoparticles;
- wherein the formulation has a particle diameter in a range of 240 nm to 700 nm;
- wherein the OmpA is derived from Shigella flexneri 2a 2457T;
- wherein the formulation is configured for an oral administration;
- and
- wherein the formulation has a mena particle size of 535.9 nm, a particle size distribution of 0.470 and a zeta potential of −20.78.
12 more elements
Dependencies are inferred from the claim text.