John P Hennessey

Glycoconjugate vaccines are a critical component of the medical arsenal against infectious diseases. This established field continues, however, to experience failures in the clinic. The lack of fundamental understanding of factors controlling clinical efficacy of glycoconjugate vaccines is discussed while key parameters demanding focused and collab...

Glycoconjugate vaccines have proven to be highly effective in preventing a variety of bacterial diseases. Publicly available data provides a partial understanding of best practices to design and manufacture new vaccines of this type without undue risk of technical failure (e.g. consistent quality, yield, stability, cost of goods, supply) or immunol...

Background: Recurrent vulvovaginal candidiasis (RVVC) is a problematic form of mucosal Candida infection, characterized by repeated episodes per year. Candida albicans is the most common cause of RVVC. Currently, there are no immunotherapeutic treatments for RVVC. Methods: This exploratory randomized, double-blind, placebo-controlled trial evalu...

Background Military trainees are at increased risk for Staphylococcus aureus colonization and infection. Disease prevention strategies are needed, but a S. aureus vaccine does not currently exist. Methods We enrolled US Army Infantry trainees (Fort Benning, GA) in a phase 2, randomized, double-blind, placebo-controlled trial of NDV-3A, a vaccine c...

The structural stability of the Haemophilus influenzae type b (Hib) capsular polysaccharide, polyribosylribitolphosphate (PRP) in an aluminum hydroxide adsorbed, polysaccharide-protein conjugate vaccine was monitored using modifications of an HPLC assay developed by Tsai et al. [Tsai C-M, Gu X-X, Byrd RA. Quantification of polysaccharide in Haemoph...

A Phase 1b/2a clinical trial of NDV-3A vaccine containing a Candida albicans recombinant Als3 protein formulated with alum protected women <40 years old from recurrent vulvovaginal candidiasis (RVVC). We investigated the potential use of anti-Als3p sera as surrogate marker of NDV-3A efficacy. Pre- and post-vaccination sera from subjects who experie...

A Phase 1b/2a clinical trial of NDV-3A vaccine containing a Candida albicans recombinant Als3 protein protected women <40 years old from recurrent vulvovaginal candidiasis (RVVC). We investigated the potential use of anti-Als3p sera as surrogate marker of NDV-3A efficacy. Pre- and post-vaccination sera from subjects who experienced recurrence of VV...

Background NDV-3A is a vaccine containing a recombinant version of the Candida albicans Als3 protein (Als3p) formulated with aluminum hydroxide (AlOH). A recently completed exploratory Phase 2a clinical trial of NDV-3A, referred to as NDV3A-003, showed that women with recurrent vulvovaginal candidiasis (RVVC) could be protected from recurrence of V...

Safe and efficacious vaccines are arguably the most successful medical interventions of all time. Yet the ongoing discovery of new pathogens, along with emergence of antibiotic-resistant pathogens and a burgeoning population at risk of such infections, imposes unprecedented public health challenges. To meet these challenges, innovative strategies t...

Currently licensed vaccines have emanated from a relatively small number of production platforms. As shown in Table 9.1, these platforms have produced a large number of licensed products and are currently being used to develop a significant number of new vaccines that are currently under clinical evaluation.

Staphylococcus aureus is the leading cause of skin and skin structure infections (SSSI) in humans. Moreover, the high frequency of recurring SSSI due to S. aureus , particularly methicillin-resistant strains (MRSA), suggests infection induces suboptimal anamnestic defenses. The present study addresses the hypothesis that interleukin-17A (IL-17A) an...

Multiple-antigen vaccines are an increasing focus of biotherapeutic development targeting infectious disease. Three recombinant antigens, based on Candida albicans proteins, have independently induced protection in animal models of candidal infections; agglutinin-like sequence 3 (rAls3), hyphally regulated protein 1 (rHyr1), and secreted aspartyl p...

Significance Staphylococcus aureus is an opportunistic pathogen of the normal human flora. It is among the most frequent causes of cutaneous abscesses, leading to life-threatening invasive infection. Incomplete understanding of host defenses against S. aureus skin or invasive infection has hindered development of effective vaccines to address these...

Recent perspectives forecast a new paradigm for future “third generation” vaccines based on commonalities found in diverse pathogens or convergent immune defenses to such pathogens. For Staphylococcus aureus, recurring infections and a limited success of vaccines containing S. aureus antigens imply that native antigens induce immune responses insuf...

We have previously reported that vaccination with rAls3p-N protein of Candida albicans, formulated with alum adjuvant (also designated as NDV-3) protects immunocompetent mice from, lethal disseminated candidiasis and mucosal oropharyngeal candidiasis. NDV-3 vaccine was recently, tested in a Phase 1 clinical trial and found to be safe, well-tolerate...

The investigational vaccine, NDV-3, contains the N-terminal portion of the Candida albicans agglutinin-like sequence 3 protein (Als3p) formulated with an aluminum hydroxide adjuvant in phosphate-buffered saline. Preclinical studies demonstrated that the Als3p vaccine antigen protects mice from oropharyngeal, vaginal and intravenous challenge with C...

A real-time quantitative polymerase chain reaction (PCR)-based method was developed to measure the concentration of recombinant adenoviral vector genomes in purified virus bulks and final container samples of monovalent and multivalent human immunodeficiency virus (HIV) adenoviral vector vaccine candidates. This method, referred to as the genome qu...

A sensitive and reproducible method to determine the in vitro infectious potency of a pentavalent reassortant rotavirus vaccine (RotaTeq) has been developed as an alternative to classical potency assays. Potency was determined based on cell-based viral replication followed by quantitative reverse-transcription polymerase chain reaction (RT-QPCR) an...

An approach was developed to align release and end-expiry specifications for molecular size for the polyvalent pneumococcal polysaccharide vaccine (PNEUMOVAX 23). Each of the 23 polysaccharide components of the vaccine was separately subjected to ultrasonication to produce a series of preparations of decreasing weight-average molecular mass (Mw). T...

PedvaxHIB is a pediatric vaccine that protects children from severe disease caused by the gram-negative bacterium Haemophilus influenzae type b (Hib). The vaccine is made by chemically conjugating Hib capsular polysaccharide to the outer membrane protein complex of Neisseria meningitidis. The protein-conjugated vaccine has proven to be extremely ef...

Purified capsular polysaccharide preparations from Streptococcus pneumoniae that are used for vaccine production typically contain residual levels of C-polysaccharide (C-Ps). Residual C-Ps is typically found in one of two forms, either chemically linked to the capsular polysaccharide (bound) or present by itself (free). Two analytical methods have...

In accordance with the US Code of Federal Regulations 21CFR 211.84 (6)(d)(1), a specific identity test must be performed for the release of chromatography media (stationary phase) before use in production of human pharmaceuticals. Due to the complexity of the physical and chemical properties of these media, i.e., variable particle morphology, insol...

A simple, accurate, and precise method to determine adenovirus particle concentration using 260-nm absorbance was developed as an enhancement to the method of Maizel et al. (1968, Virology 36, 115-125). This modified method ensures complete disruption of virus particles and viral DNA prior to absorbance measurements, therefore eliminating absorbanc...

A method utilizing NMR spectroscopy has been developed to confirm the identity of bacterial polysaccharides used to formulate a polyvalent pneumococcal polysaccharide vaccine. The method is based on 600 MHz proton NMR spectra of individual serotype-specific polysaccharides. A portion of the anomeric region of each spectrum (5.89 to 4.64 ppm) is com...

A technique using high performance size exclusion chromatography (HPSEC) with rate nephelometry (RN) detection has been developed to simultaneously measure the relative molecular size and antigenicity of the bacterial polysaccharide components in a multivalent pneumococcal vaccine, PNEUMOVAX 23. This assay was used to establish stability profiles f...

Manufacture of VAQTA, an inactivated hepatitis A vaccine, uses state-of-the-art technologies in cell culture and bioprocessing science, which have made it possible to routinely produce the vaccine at manufacturing scale. VAQTA consists of an attenuated strain of hepatitis A virus that is highly purified and formaldehyde-inactivated, then formulated...

Manufacture of VAQTA, an inactivated hepatitis A virus vaccine, includes extensive purification of the intact virus particle to remove endogenous components from the host cell culture lysate as well as compounds introduced in the upstream purification process. Analysis of the final purified hepatitis A virus product by SDS-PAGE prior to inactivatio...

We conducted a multicenter, single-blind, randomized comparisons of the immunogenicity and safety of three manufacturing-scale lots of 7.5 micrograms liquid Haemophilus influenzae type b polysaccharide- Neisseria meningitidis conjugate vaccine (PRP-OMPC) and a single lot of 15.0 micrograms lyophilized PRP OMPC. A total of 908 infants were entered i...

We conducted a multicenter, single-blind, randomized comparison of the immunogenicity and safety of three manufacturing-scale lots of 7.5 μg liquid Haemophilus influenzae type b polysaccharide-Neisseria meningitidis conjugate vaccine (PRP-OMPC) and a single lot of 15.0 μg lyophilized PRP-OMPC. A total of 908 infants were entered into the study. Eac...

Purified capsular polysaccharide preparations from Streptococcus pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were analyzed by high performance size exclusion chromatography (HPSEC) with multi-angle laser light scattering (MALLS), specific viscosity (SV), and refractive index (RI) detection to determine the molecular size and molar mass of...

The molar mass and molar mass distribution of polyribosyl ribitol phosphate (PRP) was determined using high performance size-exclusion chromatography with light scattering, viscometric, and refractive index detection. Light scattering data showed the PRP preparations to have an Mw of ca. 75,000, and a polydispersity (Mw/Mn) of about 1.46. Based on...

The capsular polysaccharide from Haemophilus influenzae type b (polyribosyl ribitol-phosphate; PRP) and the capsular polysaccharides from Streptococcus pneumoniae types 6B, 14, 18C, and 23F (Pn6B, Pn14, Pn18C, and Pn23F) were subjected to acid hydrolysis using hydrofluoric (HF) and/or trifluoroacetic acid (TFA) and high-pH anion-exchange chromatogr...

Microcarrier culture was investigated for the propagation of attenuated hepatitis A vaccine in the anchorage-dependent human fibroblast cell line, MRC-5. Cells were cultivated at 37 degrees C for one to two weeks, while virus accumulation was performed at 32 degrees C over 21 to 28 days. The major development focus for the microcarrier process was...

Reconstituted proteoliposomes containing functional Neurospora crassa plasma membrane H(+)-ATPase molecules oriented predominantly with their cytoplasmic surface exposed were treated with trypsin and then subjected to Sepharose CL-6B column chromatography to remove the liberated peptides. The peptides remaining associated with the liposomes were th...

The transmembrane topography of the Neurospora crassa plasma membrane H(+)-ATPase has been investigated using purified, reconstituted components and direct protein chemical techniques. Reconstituted proteoliposomes containing H(+)-ATPase molecules oriented predominantly with their cytoplasmic surface facing outward were treated with trypsin to libe...

Reconstituted proteoliposomes containing Neurospora plasma membrane H+-ATPase molecules oriented predominantly with their cytoplasmic portion facing outward have been used to determine the location of the NH2 and COOH termini of the H+-ATPase relative to the lipid bilayer. Treatment of the proteoliposomes with trypsin in the presence of the H+-ATPa...

A procedure for successful analysis of the hydrophobic tryptic peptides of the Neurospora crassa plasma membrane H+-ATPase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is described. The features of this procedure that are essential for the best results include (i) treatment of the hydrophobic peptide samples with neat tri...

A highly effective procedure for fragmenting the Neurospora crassa plasma membrane H+-ATPase and purifying the resulting peptides is described. The enzyme is cleaved with trypsin to form a limit digest containing both hydrophobic and hydrophilic peptides, and the hydrophobic and hydrophilic peptides are then separated by extraction with an aqueous...

In a previous communication, a water-soluble, hexameric form of the Neurospora crassa plasma membrane H+-ATPase was described (Chadwick, C. C., Goormaghtigh, E., and Scarborough, G. A. (1987) Arch. Biochem. Biophys. 252, 348-356). To facilitate physical studies of the hexamers, the H+-ATPase isolation procedure has been improved, resulting in a str...

Cytoplasmic malate dehydrogenase [L)-malate:NAD+ oxidoreductase, EC 1.1.1.37) and glyceraldehyde-3-phosphate dehydrogenase (D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase, EC 1.2.1.12) homologues from two shallow-living and three deep-living fishes were examined for the effects of hydrostatic pressure on enzyme activity and susceptibility to inac...

CD measurements were made for calmodulin and its calcium (Ca2+) complexes at different ionic strengths and Ca2+ concentrations. Calmodulin at an ionic strength of 0.00M and in the absence of Ca2+ exists as an α-helical protein with a negligible amount of β-sheet. An increase in ionic strength, whether or not Ca2+ is present, increases α-helix at th...

The inactivation by hydrostatic pressure of muscle-type lactate dehydrogenase (M4-LDH, EC 1.1.1.27; L-lactate: NAD+ oxidoreductase) homologues from five shallow-living and six deep-living marine teleost fishes was compared. The pressures which inactivate these enzymes are much higher than the pressures experienced by any of the species. To determin...

The susceptibility to inactivation by hydrostatic pressure of the tetrameric (Fig. 1) muscletype (M4) lactate dehydrogenase homologues (LDH, EC 1.1.1.27;l-lactate: NAD+ oxidoreductase) from six confamilial macrourid fishes was compared at 4 °C. These marine teleost fishes occur over depths of 260 to 4815 m. The pressures necessary to half-inactivat...

Piperitone was isolated from a sample of commercial oil of peppermint (Mentha piperita) using gentle procedures to avoid racemization. Comparison of its CD spectrum with the spectra of piperitone standards indicates that the isolated peppermint piperitone is 76–85 % the (+)-enantiomer. Piperitone as it occurs in Mentha piperita is thus (+)-4S-piper...

Seven types of error that may interfere with the analysis of protein circular dichroism (CD) spectra for secondary structure are examined. Three of these errors are operational encompassing wavelength synchronization, and proper choice of spectral bandwidth and scan speed. Three are experimental involving intensity adjustments and two sources of ba...

A change in the secondary structure of transcarboxylase from quaternary interactions is monitored by circular dichroism spectroscopy. The change is traced to interactions among the six polypeptides that make up the 12SH subunit. It is fully reversible and is not a result of the conditions used to dissociate the enzyme. Our new method of analyzing c...

A method is presented for predicting the secondary structure of a protein from its circular dichroism (CD) spectrum. Eight types of secondary structures are considered: helix; parallel and antiparallel beta strand; types I, II, and III beta turn; all other beta turns combined; and "other" structures. The method is based on mathematical calculation...

Typescript (photocopy). Thesis (Ph. D.)--Oregon State University, 1986. Includes bibliographical references (leaves 81-88).