Vaccine 2014-05-19

Quantitative high throughput analytics to support polysaccharide production process development

Aaron Noyes, Ranga Godavarti, Nigel Titchener-Hooker, Jonathan Coffman, Tarit Mukhopadhyay

Index: Vaccine 32(24) , 2819-28, (2014)

Full Text: HTML

Abstract

The rapid development of purification processes for polysaccharide vaccines is constrained by a lack of analytical tools current technologies for the measurement of polysaccharide recovery and process-related impurity clearance are complex, time-consuming, and generally not amenable to high throughput process development (HTPD). HTPD is envisioned to be central to the improvement of existing polysaccharide manufacturing processes through the identification of critical process parameters that potentially impact the quality attributes of the vaccine and to the development of de novo processes for clinical candidates, across the spectrum of downstream processing. The availability of a fast and automated analytics platform will expand the scope, robustness, and evolution of Design of Experiment (DOE) studies. This paper details recent advances in improving the speed, throughput, and success of in-process analytics at the micro-scale. Two methods, based on modifications of existing procedures, are described for the rapid measurement of polysaccharide titre in microplates without the need for heating steps. A simplification of a commercial endotoxin assay is also described that features a single measurement at room temperature. These assays, along with existing assays for protein and nucleic acids are qualified for deployment in the high throughput screening of polysaccharide feedstreams. Assay accuracy, precision, robustness, interference, and ease of use are assessed and described. In combination, these assays are capable of measuring the product concentration and impurity profile of a microplate of 96 samples in less than one day. This body of work relies on the evaluation of a combination of commercially available and clinically relevant polysaccharides to ensure maximum versatility and reactivity of the final assay suite. Together, these advancements reduce overall process time by up to 30-fold and significantly reduce sample volume over current practices. The assays help build an analytical foundation to support the advent of HTPD technology for polysaccharide vaccines. It is envisaged that this will lead to an expanded use of Quality by Design (QbD) studies in vaccine process development.


Related Compounds

Related Articles:

Escherichia coli kduD encodes an oxidoreductase that converts both sugar and steroid substrates.

2014-06-01

[Appl. Microbiol. Biotechnol. 98(12) , 5471-85, (2014)]

The N terminus of type III secretion needle protein YscF from Yersinia pestis functions to modulate innate immune responses.

2015-04-01

[Infect. Immun. 83(4) , 1507-22, (2015)]

Fingerprinting profile of polysaccharides from Lycium barbarum using multiplex approaches and chemometrics.

2015-07-01

[Int. J. Biol. Macromol. 78 , 230-7, (2015)]

Characterization of the Caenorhabditis elegans HIM-6/BLM helicase: unwinding recombination intermediates.

2014-01-01

[PLoS ONE 9(7) , e102402, (2014)]

Production of the Escherichia coli common pilus by uropathogenic E. coli is associated with adherence to HeLa and HTB-4 cells and invasion of mouse bladder urothelium.

2014-01-01

[PLoS ONE 9(7) , e101200, (2014)]

More Articles...