Tuesday, October 18, 2011

Rapid Methods Session 2

The first speaker during this session was Claudio D. Denoya, PhD, Adjunct Professor Department of Molecular and Cell Biology, University of Connecticut. Dr. Denoya discussed the current microbiology curricula taught within academic institutions and the role pharmaceutical science and modern microbiological methods should play in these programs. The focus was to understand whether microbiology students and the courses they take are appropriate in preparing individuals for a career in microbiology within the pharmaceutical industry.

When reviewing the microbiology curriculum, many courses focus on biochemistry and very few will cover the application of microbiology concepts in industry, especially pharmaceuticals. Additionally, one of the areas that are absent in the academic curricula is how to assess, validate and implement alternative, molecular and rapid microbiological methods. Because alternative methods will play an important role in the future of contamination monitoring and control, and QC microbiology activities, it is critical that these concepts be included in academic courses, especially since this enhanced knowledge and associated skill sets will be the expectation for pharma microbiologists who will want to move up the technical career ladder.

As a case study, Dr. Denoya stated that out of 65 graduates from a scientific masters program in applied genomics (at the University of Connecticut), 63 of them obtained careers in their first choice within the pharmaceutical industry. The remaining students returned to school for advanced degrees.

The next speaker was Rudolf Gilmanshin, PhD, Vice President, Advanced Platform Research, Pathogenetix, who presented on a single-molecule technology for broadband detection and identification of bacteria. The technology is based on genome sequence scanning, or GSS. Generally, genomic DNA is obtained from a sample under evaluation, and is tagged, linearized and detected in a microfluidic chip. The DNA travels through areas of focused laser light (within the microchip) and the responses are compared against an internal database.

Long fragments of DNA are required from the sample (60-350 kb or 20-115 um), must be double stranded and free of nicks, be free floating in solution, be able to hybridize with the fluorescent tags, and be of high purity. Following lysis of the sample organisms, preparation occurs in about 3 hours in an automated washing and purification instrument. The fragment is then stretched within the microfluidics chip, and will then pass through separate lasers, which will excite the fluorescent tags. Organism concentration for use in the system is between 10^7-10^9 cells.

Restriction enzymes and the tags generate fluorescent signatures used for fingerprinting. Because of the uniqueness of underlying genomic sequences, hybridized tags generate the unique signals. The system is extremely sensitive, with a detection level of 0.1% sensitivity against the internal database. Beta systems will be placed in hospital clinical labs in Q4, 2012.

The last presentation in this session was made by Alessio Fantuzzi, PhD, Microbiological Project Supervisor, and Michele Bosi, Quality Control Manager, Chiesi Pharmaceutical. They presented a case study in development and qualification of an alternative method for the release of non-sterile and sterile products (e.g., sterility testing).

For sterility testing, they perform the compendial sterility test; however, they have added an additional reading phase during the test incubation period using the ATP bioluminescent Pallchek system. The use of the Pallcheck system represents a qualitative assessment of the growth of organisms if they were present in the original sample. To demonstrate equivalence against the compendial methods, they followed the validation guidance as specified in USP 1223 and Ph. Eur. 5.1.6.

For non-sterile product the incubation period lasts for 24 hours, which is the time it takes for the system to detect the number of organisms that is at a threshold level higher than background noise. For sterile product, after a 48-hour incubation period, they filter the media and assess the media for microorganisms using the Pallchek system. To support this rapid sterility test, they also utilize an enhanced environmental monitoring and risk assessment program, and the total sterility testing program has been reduced from 14 days to 8 days.

The success of these validation programs have prompted their company to implement rapid methods ofr all of their future products/formulations.


Post a Comment