ABSTRACT
Peptides and monoclonal antibodies have both emerged as important therapeutic modalities, but each has challenges which limit their use. Non-recombinant chemical conjugation of peptides onto antibodies has the potential to minimize or eliminate altogether many of these limitations. Once such approach, pioneered by CovX has created the possibility for rapid stoichiometric fusion of pharmacophores to a single antibody platform. These molecules, called CovX-Bodies, maintain both the pharmacologic properties of a given peptide and the pharmacokinetic properties of a monoclonal antibody. The result is a new class of molecules wherein each component contributes desirable traits. In this paper, we demonstrate the use of immunoassay and two-dimensional liquid chromatography mass spectrometry (2DLC/MS) in combination to investigate the antibody conjugates of Glucagon-like peptide-1 (GLP-1) and analogs for intact protein metabolite identification directly from mouse serum. The information gained from combining these approaches has helped guide and expedite the optimization of our drug product development efforts.
Subject(s)
Chromatography, High Pressure Liquid/methods , Enzyme-Linked Immunosorbent Assay/methods , Glucagon-Like Peptide 1/blood , Mass Spectrometry/methods , Peptides/blood , Venoms/blood , Amino Acid Sequence , Animals , Antibodies, Anti-Idiotypic/immunology , Antibody Specificity , Chromatography, Affinity , Exenatide , Male , Mice , Molecular Sequence DataABSTRACT
Measles vaccination via the aerosol route has proven effective under field conditions, using vaccine reconstituted prior to nebulization. Inhalation of a dry powder aerosol vaccine would have additional benefits, including easier logistics of administration, reduced cold chain dependence and the potential of single dose administration. We have evaluated two candidate dry powder measles vaccine formulations in macaques. Specific immune responses were demonstrated, but levels of immunity were lower than in animals vaccinated by injection or by nebulized aerosol. These studies provide proof of principle that dry powder inhalation is a possible route for measles vaccination, but suggest that either the vaccine formulation or the method of delivery need to be improved for a better immune response.
Subject(s)
Measles Vaccine/administration & dosage , Measles Vaccine/therapeutic use , Administration, Inhalation , Anesthesia, Endotracheal , Animals , Antibodies, Viral/analysis , Antibodies, Viral/biosynthesis , Injections , Lung/metabolism , Macaca fascicularis , Measles/immunology , Measles/prevention & control , Measles virus/chemistry , Neutralization Tests , Particle Size , Powders , RNA, Viral/analysis , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction , Vaccination/methods , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/therapeutic useABSTRACT
PURPOSE: To evaluate an anomalous increase in the specific surface area of budesonide during storage postmicronization. METHODS: Budesonide was micronized using a conventional air-jet mill. Surface areas and total pore volumes were measured using nitrogen sorption. Porosity was measured using mercury intrusion porosimetry. Particle size was measured using laser diffraction. RESULTS: Budesonide exhibited a surface area increase of 22 +/- 2% when stored at 25 degrees C following micronization. The rate of surface area increase was lower at 20 degrees C. suggesting a temperature-dependent stress relaxation mechanism for the micronized particles. The increase in surface area was accompanied by: (a) an increase in total pore volume: (b) a shift of the pore size distribution to smaller pore sizes; (c) a decrease in size of particles above approximately 1 microm; and (d) an increase in rugosity/surface roughness. CONCLUSIONS: Freshly micronized budesonide exhibited an unusual and significant postmicronization increase in specific surface area upon storage under ambient conditions. Postmicronization stress-relief by intraparticle crack formation, crack propagation with time, and particle fracture seems to be the primary mechanism behind this surface area increase.
