The bioanalysis of plasma samples generated from in vivo studies of therapeutic proteins is of increasing interesting in the biopharmaceutical industry. The conventional ELISA approach has a long assay development time which can limit use in the early discovery and development of protein-based drugs. In this study, an LC−MS/MS bioassay was developed for the quantification of somatropin and a therapeutic human monoclonal antibody. The assay used bovine fetuin as an internal standard and a two-dimensional solid-phase extraction for the cleanup of the plasma digest. Sample extracts were resolved on an analytical size column using a 6 min LC gradient and analyzed using a triple-quadruple mass spectrometer. The linearity of the assay for somatropin was established from 1 to 1000 μg/mL with accuracy and precision within 15%. This LC−MS approach was also applied to a rat pharmacokinetic study of the therapeutic monoclonal antibody with a lower quantitation limit of 0.5 μg/mL. The LC−MS assay had improved accurac...

LC−MS/MS Approach for Quantification of Therapeutic Proteins in Plasma Using a Protein Internal Standard and 2D-Solid-Phase Extraction Cleanup

B. anthracis, the causative agent for anthrax, has been well studied for over 150 years. Due to the genetic similarities among various Bacillus species, as well as its existence in both a spore form and a vegetative state, the detection and specific identification of B. anthracis have been proven to require complex techniques and/or laborious methods. With the heightened interest in the organism as a potential biological threat agent, a large number of interesting detection technologies have recently been developed, including methods involving immunological and nucleic acid-based assay formats. The technologies range from culture-based methods to portable Total Analysis Systems based on real-time PCR. This review with 170 references provides a brief background on the toxicology and epidemiology of B. anthracis, discusses challenges associated with its detection related to genetic similarities to other species, and reviews immunological and, with greater emphasis, nucleic acid-based detection systems.

Bacillus anthracis: toxicology, epidemiology and current rapid-detection methods.

Free-living microbes are provided in which the nucleic acid has been modified so that the microbe is attenuated for proliferation and/or which comprise genetic mutations that attenuate the ability of the microbe to repair its nucleic acid Methods of using the modified microbes for the loading, activation, and/or maturation of antigen-presenting cells are also provided Vaccine compositions comprising the modified microbes and/or the antigen-presenting cells and methods of using the vaccines are also provided The microbes may be further modified to include heterologous antigens, such as tumor antigens or infectious disease antigens, for use as a vaccine against cancer or infectious diseases

Modified free-living microbes, vaccine compositions and methods of use thereof

Live attenuated and inactivated viral vaccine formulation and nasal delivery: Potential and challenges

The current human anthrax vaccines licensed in the US and UK consist of aluminum hydroxide-adsorbed or alum-precipitated culture supernatant material from fermentor cultures of toxigenic noncapsulated strains of Bacillus anthracis. The threat of B. anthracis being used as a biowarfare agent has led to a wider usage of these vaccines, which has heightened concerns regarding the need for frequent boosters and the occasional local reactogenicity associated with vaccination. These concerns have provided the impetus for the development of better characterized vaccines. This review summarizes the work of numerous laboratories in the search for alternative vaccines against anthrax that are well tolerated, provide long-lasting immunity, and are efficacious.

Henry Hsu

Papers

Validation of an anti-PA-ELISA for the potency testing of anthrax vaccine in mice.

Target alternative vaccine safety testing strategies for pertussis toxin