IP Library › Granted Patent US 12,303,566
Granted Patent B2
US 12,303,566 · App. 17/193,999 · Granted May 20, 2025

Immunostimulatory compositions and methods of use thereof

Inventors: Darrell Irvine (Arlington, MA); Haipeng Liu (Malden, MA)
Assignee: Massachusetts Institute of Technology
A61K47/645A61K39/0005A61K39/0011A61K39/12A61K39/21A61K39/39A61K47/542A61K47/543A61K47/643A61K47/6455
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Quick Facts
Patent No.
US 12,303,566
App. No.
17/193,999
Granted
May 20, 2025
Kind
B2
Abstract

Lipid conjugates for enhanced delivery of cargo to the lymph nodes are disclosed. The lipid conjugates typically include three domains: a lipophilic domain that binds to albumin, a polar block domain, and a cargo such as a molecular adjuvant or immunostimulatory compound (such as an oligonucleotide) or antigenic peptide. Depending on the cargo, the length and compositions of the polar block can be tailored to push the equilibrium toward albumin binding, stable micelle formation, or cell insertion. The conjugates can be administered to a subject, for example, a subject with cancer or an infection, to induce or enhance a robust immune response in the subject.

Claims (27)

1. An amphiphilic albumin-binding conjugate comprising

(a) a lipid component;

(b) an optional polar component; and

(c) a ligand for a pattern recognition receptor;

wherein the ligand for a pattern recognition receptor is bound directly to the lipid or is bound to the lipid via a linker,

wherein the conjugate is sufficiently soluble that the lipid binds to albumin under physiological conditions, and

wherein a plurality of the conjugates can spontaneously form micelles in aqueous solution.

2. The conjugate of claim 1 , wherein the ligand for the pattern recognition receptor is an immunostimulatory oligonucleotide.

3. The conjugate of claim 2 , wherein the immunostimulatory oligonucleotide is sufficiently polar to reduce the ability of the lipid to insert into a cellular plasma membrane.

4. The conjugate of claim 1 , wherein the pattern recognition receptor is a retinoic acid-inducible gene I (RIG-I)-like receptor.

5. The conjugate of claim 4 , wherein the RIG-I-like receptor is RIG-I or melanoma differentiation-associate gene 5 (MDAS).

6. The conjugate of claim 1 wherein the ligand for a pattern recognition receptor is bound to the lipid via a linker.

7. The conjugate of claim 6 wherein the linker is an oligonucleotide linker.

8. The conjugate of claim 7 wherein the oligonucleotide linkers comprises “N” consecutive guanines, wherein N is between 0-2 or is between 3-10.

9. The conjugate of claim 8 comprising the structure L-5′-G n -ON-3′, wherein “L” the lipid, “G” is a guanine, “n” is 0-2, and “ON” is an immunostimulatory oligonucleotide.

10. The conjugate of claim 1 wherein the conjugate exhibits increased accumulation in the lymph node when administered to a subject in vivo compared to administration of the ligand for a pattern recognition receptor alone.

11. The conjugate of claim 1 wherein the lipid is a diacyl lipid.

12. The conjugate of claim 11 wherein the acyl chains of the lipid comprise 12-30 hydrocarbon units.

13. The conjugate of claim 1 wherein the pattern recognition receptor is a Toll-like receptor.

14. The conjugate of claim 2 wherein the immunostimulatory oligonucleotide comprises CpG.

15. The conjugate of claim 2 wherein the immunostimulatory oligonucleotide has a phosphorothioate (PS) backbone.

16. The conjugate of claim 2 wherein the immunostimulatory oligonucleotide comprises 20 or more nucleic acids.

17. A vaccine adjuvant comprising a plurality of the amphiphilic albumin-binding conjugates of claim 1 .

18. The conjugate of claim 6 wherein the linker is selected from the group consisting of hydrophilic polymers, a string of hydrophilic amino acids, polysaccharides or a combination thereof.

19. The conjugate of claim 18 wherein the linker comprises “N” consecutive polyethylene glycol units, wherein N is between 25-50.

20. The conjugate of claim 13 , wherein the Toll-like Receptor is Toll-like receptor 9, Toll-like receptor 7, or Toll-like receptor 3.

21. The conjugate of claim 20 , wherein the Toll-like receptor is Toll-like receptor 9.

Assignments (6)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: IRVINE, DARRELL; LIU, HAIPENG
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 057336/0440 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: IRVINE, DARRELL; MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 057336/0807 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: IRVINE, DARRELL
To: HOWARD HUGHES MEDICAL INSTITUTE
Reel/Frame 057338/0595 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: LIU, HAIPENG
Reel/Frame 057338/0702 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: LIU, HAIPENG
To: HOWARD HUGHES MEDICAL INSTITUTE
Reel/Frame 057338/0786 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 31, 2021
From: HOWARD HUGHES MEDICAL INSTITUTE
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 057339/0450 →
Continuity (5)
Continuation 16040181 · Jul 19, 2018
Continuation 14790432 · Jul 2, 2015
Division 13844075 · Mar 15, 2013
Provisional Application 61620518 · Apr 5, 2012
Related Publication 20210187112A1 · Jun 24, 2021
References Cited (148)
US 7615539B2 · Uhlmann · 2009 [cited by applicant]
US 7741297B2 · Jiang · 2010 [cited by applicant]
US 9107904B2 · Irvine · 2015 [cited by examiner]
US 9426778B2 · Onozawa · 2016 [cited by applicant]
US 10029016B2 · Irvine · 2018 [cited by examiner]
US 10953105B2 · Irvine · 2021 [cited by examiner]
US 20060246126A1 · Allen et al. · 2006 [cited by applicant]
US 20070104654A1 · Hsieh · 2007 [cited by applicant]
US 20070154398A1 · Wang · 2007 [cited by applicant]
US 20080139262A1 · Lin · 2008 [cited by applicant]
US 20100183639A1 · Uhlmann · 2010 [cited by applicant]
US 20110300163A1 · Champion · 2011 [cited by applicant]
US 20120087949A1 · Tan · 2012 [cited by applicant]
US 20120121606A1 · Ruben · 2012 [cited by applicant]
US 20120129199A1 · Daftarian · 2012 [cited by applicant]
US 20120294931A1 · Kim et al. · 2012 [cited by applicant]
US 20140099337A1 · Davis · 2014 [cited by applicant]
US 20140162944A1 · Tiberg · 2014 [cited by applicant]
US 20140255378A1 · Watkins · 2014 [cited by applicant]
US 20140294932A1 · Kim · 2014 [cited by applicant]
US 20150044208A1 · Castanheira Aires da Silva · 2015 [cited by applicant]
US 20150232514A1 · Dockal · 2015 [cited by applicant]
US 20150283205A1 · Phipps · 2015 [cited by applicant]
US 20160228573A1 · Niyikiza et al. · 2016 [cited by applicant]
US 20160361360A1 · Chang et al. · 2016 [cited by applicant]
US 20170224797A1 · Popescu et al. · 2017 [cited by applicant]
US 20180028634A1 · Chen · 2018 [cited by applicant]
US 20190062457A1 · Elias et al. · 2019 [cited by applicant]
US 20200230221A1 · Irvine et al. · 2020 [cited by applicant]
US 20200345853A1 · Irvine et al. · 2020 [cited by applicant]
US 20230372395A1 · Irvine et al. · 2023 [cited by applicant]
US 20240082373A1 · Irvine et al. · 2024 [cited by applicant]
EP 0491628 · 1992 [cited by applicant]
JP 8510255A · 1996 [cited by applicant]
JP 2010507386A · 2010 [cited by applicant]
JP 2014504294A · 2014 [cited by applicant]
JP 2015514108A · 2015 [cited by applicant]
KR 1020150030724A · 2015 [cited by applicant]
WO 94001138 · 1994 [cited by applicant]
WO 9426778 · 1994 [cited by applicant]
WO 03066649 · 2003 [cited by applicant]
WO 2004029277 · 2004 [cited by applicant]
WO 2005117984 · 2005 [cited by applicant]
WO 2008115319 · 2008 [cited by applicant]
WO 2008121949 · 2008 [cited by applicant]
WO 2008139262 · 2008 [cited by applicant]
WO 2010071852 · 2010 [cited by applicant]
WO 2012082841A2 · 2012 [cited by applicant]
WO 2013151771A1 · 2013 [cited by applicant]
WO 2014011987A1 · 2014 [cited by applicant]
WO 2014204762A1 · 2014 [cited by applicant]
WO 2015057834A1 · 2015 [cited by applicant]
WO 2016069647A1 · 2016 [cited by applicant]
WO 2017035009A1 · 2017 [cited by applicant]
WO 2017137477A1 · 2017 [cited by applicant]
WO 2017141243A1 · 2017 [cited by applicant]
WO 2019060425A1 · 2019 [cited by applicant]
U.S. Appl. No. 16/825,482 (Year: 2020). [cited by examiner]
U.S. Appl. No. 18/339,230, filed Jun. 21, 2023, Pending. [cited by applicant]
Barthelemy et al., Comprehensive analysis of the factors contributing to the stability and solubility of autonomous human VH domains. J Biol Chem. Feb. 8, 2008;283(6):3639-3654. [cited by applicant]
Beiboer et al., Guided selection of a pan carcinoma specific antibody reveals similar binding characteristics yet structural divergence between the original murine antibody and its human equivalent. J Mol Biol. Feb. 25,… [cited by applicant]
Bejestani et al., Characterization of a switchable chimeric antigen receptor platform in a pre-clinical solid tumor model. Oncoimmunology. Jun. 20, 2017;6(10):e1342909. [cited by applicant]
Cao et al., Design of Switchable Chimeric Antigen Receptor T Cells Targeting Breast Cancer. Angew Chem Int Ed Engl. Jun. 20, 2016;55(26):7520-4. [cited by applicant]
Choi et al., Predicting antibody complementarity determining region structures without classification. Mol Biosyst. Dec. 2011;7(12):3327-34. [cited by applicant]
De Genst et al., Antibody repertoire development in camelids. Dev Comp Immunol. 2006;30(1-2):187-98. [cited by applicant]
Glienke et al., Advantages and applications of CAR-expressing natural killer cells. Front Pharmacol. Feb. 12, 2015;6:21. [cited by applicant]
Griffiths et al., Human anti-self antibodies with high specificity from phage display libraries. EMBO J. Feb. 1993;12 (2):725-34. [cited by applicant]
Irvine, Abstract IA06: Enhancing combination immunotherapy and adoptive cell therapy of cancer via lymph node-targeted vaccines. Cancer Immunology Research. Sep. 2018;6(9):two pages. [cited by applicant]
Irvine, Abstract IA37: Enhancing the function of CART cells via a universal vaccine strategy. Cancer Immunology Research. Feb. 2019;7(2):two pages. [cited by applicant]
Kim et al., Redirection of genetically engineered CAR-T cells using bifunctional small molecules. J Am Chem Soc. Mar. 4, 2015;137(8):2832-5. [cited by applicant]
Klimka et al., Human anti-CD30 recombinant antibodies by guided phage antibody selection using cell panning. Br J Cancer. Jul. 2000;83(2):252-60. [cited by applicant]
Kudo et al., T lymphocytes expressing a CD16 signaling receptor exert antibody-dependent cancer cell killing. Cancer Res. Jan. 1, 2014;74(1):93-103. [cited by applicant]
Lohmueller et al., mSA2 affinity-enhanced biotin-binding CAR T cells for universal tumor targeting. Oncoimmunology. Oct. 26, 2017;7(1):e1368604. [cited by applicant]
Ma et al., Enhanced CAR-T cell activity against solid tumors by vaccine boosting through the chimeric receptor. Science. Jul. 12, 2019;365(6449):162-168. [cited by applicant]
Ma et al., Versatile strategy for controlling the specificity and activity of engineered T cells. Proc Natl Acad Sci U S A. Jan. 26, 2016;113(4):E450-8. [cited by applicant]
Malia et al., Epitope mapping and structural basis for the recognition of phosphorylated tau by the anti-tau antibody AT8. Proteins. Apr. 2016;84(4):427-34. [cited by applicant]
Rodgers et al., Switch-mediated activation and retargeting of CAR-T cells for B-cell malignancies. Proc Natl Acad Sci U S A. Jan. 26, 2016;113(4):E459-68. [cited by applicant]
Shariat et al., P5 HER2/neu-derived peptide conjugated to liposomes containing MPL adjuvant as an effective prophylactic vaccine formulation for breast cancer. Cancer Lett. Dec. 1, 2014;355(1):54-60. [cited by applicant]
Takagi et al., Chimeric cytokine receptor can transduce expansion signals in interleukin 6 receptor alpha (IL-6Ralpha)-, IL-11Ralpha-, and gp130-low to -negative primitive hematopoietic progenitors. Mol Biol Cell. Nov. … [cited by applicant]
Urbanska et al., A universal strategy for adoptive immunotherapy of cancer through use of a novel T-cell antigen receptor. Cancer Res. Apr. 1, 2012;72(7):1844-52. [cited by applicant]
Ward et al., Binding activities of a repertoire of single immunoglobulin variable domains secreted from [cited by applicant]
Watanabe et al., Conferring CAR T Cells with Resistance to TGFβ1 Using a Signal Converter. Molecular Therapy. May 2014;22(1):S298. [cited by applicant]
Wilkie et al., Selective expansion of chimeric antigen receptor-targeted T-cells with potent effector function using interleukin-4. J Biol Chem. Aug. 13, 2010;285(33):25538-44. [cited by applicant]
Aldwell, et al., “Oral delivery of lipid-encapsulated Mycobacterium bovis BCG extends survival of the bacillus in vivo and induces a long-term protective immune response against tuberculosis”, Vaccine, 24(12):2071-2078 … [cited by applicant]
Andrews, et al., “Conjugation of lipid and CpG-containing oligonucleotide yields an efficient method for liposome incorporation”, Bioconj Chem., 22(7):1279-86 (2011). [cited by applicant]
Bachmann and Jennings, “Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns”, Nat. Rev. Immunol., 10:787-96 (2010). [cited by applicant]
Bagby, et al., “Impact of molecular weight on lymphatic drainage of a biopolymer-based imaging agent”, Pharmaceutics, 4:276-95 (2012). [cited by applicant]
Ballas, et al., “Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs”, J. Immunol., 167:4878-86 (2001). [cited by applicant]
Bedoui, et al., “Cross-presentation of viral and self antigens by skin-derived CD103+ dendritic cells”, Nat. Immunol., 10:488-95 (2009). [cited by applicant]
Bourquin, et al., “Targeting CpG oligonucleotides to the lymph node by nanoparticles elicits efficient antitumoral immunity”, J. Immunol., 181:2990-8 (2008). [cited by applicant]
Cai, et al., “Lymphatic drug delivery using engineered liposomes and solid lipid nanoparticles”, Adv. Drug Delivery Rev., 63:901-8 (2011). [cited by applicant]
Cuomo, et al., “Oligonucleotides and polynucleotides condensation onto liposome surface: effects of the base and of the nucleotide length”, Colliods Surf B Biointerfaces, 104(1):239-44 (2013). [cited by applicant]
Dass, “Lipoplex-mediated delivery of nucleic acids: factors affecting in vivo transfection”, J Mol. Med., 82(9):579-91 (2004). [cited by applicant]
Davis, “G-quartets 40 years later: from 5′-GMP to molecular biology and supramolecular chemistry”, Angew. Chem. Int. Ed. Engl., 43:668-98 (2004). [cited by applicant]
Fujita , et al., “Overview and outlook of Toll-like receptor ligand-antigen conjugate vaccines”, Ther. Deliv., 3:749-60 (2012). [cited by applicant]
Goldblatt, “Conjugate vaccines”, Clin. Exp. Immunol., 119:1-3 (2000). [cited by applicant]
Heegaard, et al., “Dendrimers for vaccine and immunostimulatory uses. A review”, Bioconj Chem., 21:405-18 (2010). [cited by applicant]
Hubbel, et al., “Materials engineering for immunomodulation”, Nature, 462:449-60 (2009). [cited by applicant]
Johansena, et al., “Lympho-geographical concepts in vaccine delivery”, J. Control Rel., 148:56-62 (2010). [cited by applicant]
Kaminskas, et al., “PEGylation of polylysine dendrimers improves absorption and lymphatic targeting following SC administration in rats”, J. Control Release, 140:108-16 (2009). [cited by applicant]
Kaminskas, et al., “Targeting the lymphatics using dendritic polymers (dendrimers)”, Adv. Drug Deliv. Rev., 63:890-900 (2011). [cited by applicant]
Keler, et al., “Antibody-targeted vaccines”, Oncogene, 26:3758-67 (2007). [cited by applicant]
Klinman, “Immunotherapeutic uses of CpG oligodeoxynucleotides”, Nat. Rev. Immunol., 4:249-59 (2004). [cited by applicant]
Kobayashi, et al., “Delivery of gadolinium-labeled nanoparticles to the sentinel lymph node: comparison of the sentinel node visualization and estimations of intra-nodal gadolinium concentration by the magnetic resonanc… [cited by applicant]
Kobayashi, et al., “Multi-Modal Nano-Probes for Radionuclide and 5-color Near Infrared Optical Lymphatic Imaging”, ACS Nano., 1:258-64 (2007). [cited by applicant]
Kwong, et al., “Induction of potent anti-tumor responses while eliminating systemic side effects via liposome-anchored combinatorial immunotherapy”, Biomaterials, 32:5134-47 (2011). [cited by applicant]
Liu, et al., “Membrane anchored Immunostimulatory Oligonucleotides for In Vivo Cell Modification and Loacalized Immunotherapy”, J. agnew. Chem. Int. Ed., 50(31):7052-7055(2011). [cited by applicant]
Liu, et al., “Structure-based programming of lymph-node targeting in molecular vaccies”, Nature, 507(7943):519-522 (2014). [cited by applicant]
Liu, et al., “DNS-based miscelles: Synthesis, micellar properties and size-dependent cel permeability”, Chem, 16(12):3791-7 (2010). [cited by applicant]
Liu, et al., “Guiding principles in the design of molecular bioconjugates for vaccine applications”, Bioconjug. Chem., 26(5):791-801 (2015). [cited by applicant]
Liu, et al., “Membrane anchored immunostimulatory oligonucleotides for in vivo cell modification and localized immunotherapy”, Angew. Chem. Int. Ed., 50(31):7052-5, Supporting Information, 8 pages (2011). [cited by applicant]
Manolova, et al., “Nanoparticles target distinct dendritic cell populations according to their size”, Eru. J. Immunol., 38:1404-13 (2008). [cited by applicant]
McLennan, et al., “Subcutaneous drug delivery and the role of the lymphatics”, Drug Discovery Today: Technol, ;2:89-96 (2005). [cited by applicant]
Mishra, et al., “Improved leishmanicidal effect of phosphorotioate antisense oligonucleotides by LDL-mediated delivery”, Biochim. Biophysica Acta, 1264(2):229-37 (1995). [cited by applicant]
Moon, et al., “Engineering nano- and microparticles to tune immunity”, Adv. Mater., 24:3724-46 (2012). [cited by applicant]
Moyle, et al., “Modern Subunit Vaccines: Development, Components, and Research Opportunities”, ChemMedChem., 8:360-76 (2013). [cited by applicant]
Oyewumi, et al., “Nano-microparticles as immune adjuvants: correlating particle sizes and the resultant immune responses”, Expert Rev. Vaccines, 9:1095-107 (2010). [cited by applicant]
Pal and Ramsey, “The role of the lymphatic system in vaccine trafficking and immune response”, Adv. Drug Delivery Rev., 63:909-22 (2011). [cited by applicant]
Pape, et al., “The humoral immune response is initiated in lymph nodes by B cells that acquire soluble antigen directly in the follicles.”, Immunity, 26(4):491-502 (2007). [cited by applicant]
Paramasivan, et al., “Circular dichroism of quadruplex DNAs: applications to structure, cation effects and ligand binding”, Methods, 43:324-31 (2007). [cited by applicant]
Perrie, et al., “Vaccine adjuvant systems: enhancing the efficacy of sub-unit protein antigens”, Int. J. Pharm., 364:272-80 (2008). [cited by applicant]
Pilishvili, et al., “Sustained reductions in invasive pneumococcal disease in the era of conjugate vaccine”, J. Infec. Dis., 201:32-41 (2010). [cited by applicant]
Rao, et al., “Biodegradable PLGA based nanoparticles for sustained regional lymphatic drug delivery”, J. Pharm. Sci., 99:2018-31 (2010). [cited by applicant]
Reddy, et al., “Exploiting lymphatic transport and complement activation in nanoparticle vaccines”, Nat. Biotechnol., 25:1159-64 (2007). [cited by applicant]
Robbins, et al., “Polysaccharide-ptotein conjugates: a new generation of vaccines”,. J. Infect. Dis., 161:821-32 (1990). [cited by applicant]
Schnorrer, et al., “The dominant role of CD8+ dendritic cells in cross-presentation is not dictated by antigen capture”, PNAS, 103:10729-34 (2006). [cited by applicant]
Senti, et al., “Intralymphatic immunotherapy”, Curr. Opin. Allergy Clin. Immunol., 9:537-43 (2009). [cited by applicant]
Singh, et al., “Nanoparticles and microparticles as vaccine-delivery systems”, Exp RevVaccine, 6(5):797-808 (2007). [cited by applicant]
Slovin, et al., “Carbohydrate vaccines as immunotherapy for cancer”, Immunol. Cell Biol., 83:418-28 (2005). [cited by applicant]
Smith, et al., “Cutting edge: conventional CD8 alpha+ dendritic cells are preferentially involved in CTL priming after footpad infection with herpes simplex virus-1”, J. Immunol., 170:4437-40 (2003). [cited by applicant]
St. John, et al., “Synthetic mast-cell granules as adjuvants to promote and polarize immunity in lymph nodes”, Nature Materials, 11:250-7 (2012). [cited by applicant]
Storhoff, et al., “Onepot colorimetric differentiation of polynucleotides with single base imperfections using gold nanoparticle probes”, J Am. Chem. Soc., 120:1959-64 (1999). [cited by applicant]
Tacken, et al., “Dendritic-cell immunotherapy: from ex vivo loading to in vivo targeting”, Nat. Rev. Immunol., 10:790-802 (2007). [cited by applicant]
Takakura, et al., “Development of a novel polymeric prodrug of mitomycin C, mitomycin C-dextran conjugate with anionic charge. II. Disposition and pharmacokinetics following intravenous and intramuscular administration”… [cited by applicant]
Tenbusch, et al., “Immunogenicity of DNA vaccines encoding simian immunodeficiency virus antigen targeted to dendritic cells in rhesus macaques”, PLoS ONE, 7:e39038 (2012). [cited by applicant]
Tsopelas, et al., “Why certain dyes are useful for localizing the sentinel lymph node”, J. Nucl. Med., 43:1377-82 (2002). [cited by applicant]
Vollmer and Krieg, “Immunotherapeutic applications of CpG oligodeoxynucleotide TLR9 agonists”, Adv. Drug Delivery Rev., 61:195-204 (2009). [cited by applicant]
Von Beust, et al., “Improving the therapeutic index of CpG oligodeoxynucleotides by intralymphatic administration”, Eur. J. Immunol., 35:1869-76 (2005). [cited by applicant]
Wilson, et al., “Lipid-based delivery of CpG oligonucleotides enhances immunotherapeutic efficacy”, Adv. Drug Deliv. Rev., 61(3):233-42 (2009). [cited by applicant]
Wong, et al., “Lymphatic drainage of skin to a sentinel lymph node in a feline model”, Ann. Surg., 214:637-40 (1991). [cited by applicant]
Wu, et al., “Fluorescence imaging of the lymph node uptake of proteins in mice after subcutaneous injection: molecular weight dependence”, Pharm. Res., 29:1843-53 (2012). [cited by applicant]
Zepp, “Principles of vaccine design—Lessons from nature”, Vaccine, 28S:C14-C24 (2010). [cited by applicant]
U.S. Appl. No. 16/644,893, filed Mar. 5, 2020, 2020-0230221, Abandoned. [cited by applicant]
U.S. Appl. No. 18/339,230, filed Jun. 21, 2023, 2024-0082373, Published. [cited by applicant]
U.S. Appl. No. 13/844,075, filed Mar. 15, 2013, U.S. Pat. No. 9,107,904, Issued. [cited by applicant]
U.S. Appl. No. 14/790,432, filed Jul. 2, 2015, U.S. Pat. No. 10,029,016, Issued. [cited by applicant]
U.S. Appl. No. 16/040,181, filed Jul. 19, 2018, U.S. Pat. No. 10,953,105, Issued. [cited by applicant]
U.S. Appl. No. 16/825,482, filed Mar. 20, 2020, 2020-0345853, Published. [cited by applicant]