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Protein matrix vaccines and methods of making and administering such vaccines

A protein and carrier protein technology, applied in the field of vaccine compositions, can solve the problems of little immune response stimulation, no stimulation, complicated efforts, etc.

Active Publication Date: 2009-10-07
PRESIDENT & FELLOWS OF HARVARD COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Many antigens, especially those associated with the pathogen capsule, stimulate little or no immune response and complicate efforts to generate effective vaccines against those antigens

Method used

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  • Protein matrix vaccines and methods of making and administering such vaccines
  • Protein matrix vaccines and methods of making and administering such vaccines
  • Protein matrix vaccines and methods of making and administering such vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0188] Example 1. Vaccine and control formulations

[0189] Capsular poly-γ-D-glutamic acid (PGA) was purchased from Vedan (Taiwan) or purified by the method of Rhie et al. (Proc. Natl. Acad. Sci. USA 100: 10925-10930, 2003). Dominant negative mutant (DNI) is a mutant form of Bacillus anthracis (B. anthracis) protective antigen (PA), prepared from Escherichia coli (Escherichia coli) by the method of Benson et al. (Biochemistry 37:3941-3948, 1998) have to. PGA and DNI proteins were extensively dialyzed against 0.05M, pH 7.4 sodium phosphate buffer (SP7.4) before use. The concentration of DNI stock solution is 30mg / mL. The PGA stock solution concentration was 134 mg / mL. Linker glutaraldehyde was purchased from Pierce as a 25% stock solution. Protein capsular matrix vaccine (PCMV) and controls were assembled in the reactions in Table 1.

[0190] Table 1. Assembly of reactions used to prepare PCMV formulations 1-3 and controls 4 and 5

[0191] Reaction #DNI PGA dH 2 O 25% g...

Embodiment 2

[0210] Example 2. Immunization and analysis of anti-DNI and anti-PGA immune responses

[0211] The soluble products of the five reactions described in Table 1 were adjusted to the same protein concentration (based on their absorbance at 280 nm). BALB / c mice approximately 5-7 weeks old from Charles River were used for figure 2 All immunization studies described. Mice were immunized with PCMV vaccines 1-3 and antigen preparation controls 4 and 5 at a dose of 20 μg of DNI protein by intraperitoneal injection on day 0. All mice were bled on day 7 and then boosted with the same dose of antigen preparation on day 10. Mice were bled again on day 17 and boosted again on day 20. Mice were bled again on day 30 and sacrificed this time. Serum was collected from blood samples after clotting and stored at -20°C. The levels of anti-PGA serum antibodies and anti-DNI serum antibodies were analyzed using enzyme-linked immunosorbent assay (ELISA). Briefly, Immulon 2HB ELISA (VWR) microti...

Embodiment 3

[0215] Example 3. Preparation and Characterization of Additional PCMVs

[0216] PCMV technology can be applied to capsular antigens of various structures and ionic charges. 23 types of Streptococcus pneumonia PS were purchased from American Type Culture Collection (ATCC) and manufactured by Merck, Inc. These PSs vary widely in their molecular structures, including strongly anionic, partially cationic, neutrally charged, phosphorylated, linear, branched, and modified in various other ways. In a pilot experiment, a subset of these PSs (4, 6B, 9V, 14, 18C, 19F, and 23F) corresponding to the seven capsular types in the Wyeth product Prevnar were taken and determined to induce IL-6 production by mouse macrophages Ability. Type 4 PS was active in the assay; lipopolysaccharide (LPS) was the control for TLR agonists. Other PS (eg, type 3), PGA and O antigen PS from F. tularensis, as well as PCMV vaccines prepared from PGA-DNI and no cross-linked controls were also assayed. This ex...

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Abstract

The invention relates to vaccine compositions having a carrier protein and an antigen of interest entrapped in a complex, methods of making such vaccines, and methods of vaccine administration.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application Serial No. 60 / 835,944, filed August 7, 2006, and US Provisional Application Serial No. 60 / 933,764, filed June 8, 2007, the specifications of which are incorporated herein by reference. [0003] Statement Regarding Federally Funded Research [0004] This invention was made with government support under Grant No U54AI057159 awarded by the National Institutes of Health (NIH). The government has certain rights in this invention. Background of the invention [0005] The present invention relates to vaccine compositions, methods of preparing vaccines and methods of administering vaccines. [0006] Many antigens, particularly those associated with the pathogen capsule, stimulate little or no immune response and complicate efforts to generate effective vaccines against those antigens. The capsule is the surface component of microorganisms, usually composed of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/385A61K39/116A61K39/02A61K39/09A61K39/095A61K31/70
CPCY02A50/30
Inventor J·J·梅卡拉诺斯
Owner PRESIDENT & FELLOWS OF HARVARD COLLEGE
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