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In the last years the LAL test has become the preferred system to test for endotoxins – for the in-process control as well as in the final inspection – and it is anchored in the pharmacopoeias. However, in the recent past, the problem of low endotoxin recovery employs the pharmaceutical microbiology. Masking – or not? Evidence gaps? And how can I close them? And how to evaluate?
These are the questions the pharmaceutical microbiologists as well as those responsible for the release have to deal with.
And last but not least, how can we handle the test in daily business in a practical manner.
Endotoxin Detection Methods
Simulation of contamination in various sample matrices
Analysis of interference in affected samples
Sample treatment for demasking
Interpretation and Comparison of Results
Closing Remarks (ECA Academy)
26-27 February 2019
Speakers
John Dubczak, General Manager at Charles River
Stefan Gärtner, Labor LS
Dr Holger Grallert, Hyglos GmbH
Dr. Andreas Karst, Haemochrom Diagnostica
Dr Johannes Reich, Microcoat Biotechnologie GmbH
Kevin Williams, bioMérieux
Veronika Wills, ACC
Dr Thomas Winkler, Lonza Cologne GmbH
Learning Objectives
- How to identify Low Endotoxin Recovery (LER)
- How to Set-up hold-time studies
- Analysis of influencing factors (Sample matrices, endotoxin, temperature, detection methods, etc.)
- Understanding the driving forces of LER
- Interpretation of test results
- Dedicated sample treatment for demasking
Background
In the last years the LAL test has become the preferred system to test for endotoxins – for the in-process control as well as in the final inspection – and it is anchored in the pharmacopoeias. However, in the recent past, the problem of low endotoxin recovery employs the pharmaceutical microbiology. Masking – or not? Evidence gaps? And how can I close them? And how to evaluate?
These are the questions the pharmaceutical microbiologists as well as those responsible for the release have to deal with.
And last but not least, how can we handle the test in daily business in a practical manner.
Target Group
- Laboratory management and staff of pharmaceutical microbiology
- Microbiologists and laboratory assistants from contract laboratories
- Scientific staff from the area Endotoxin testing
Programme
Endotoxin Detection Methods
- Definition of Endotoxins
- Nature of Endotoxin
- General detection methods
- Basic reaction of Limulus-based detection methods
- Sample handling
- Construction and interpretation of standard curve
- Mechanisms of LER can be influenced by LPS stability
- Varying size and fragility of LPS aggregates can be directly correlated to presence of chelating agents and detergents
- Elucidating conformational changes, in real time, on LPS using HSAFM
- Postitive Product Control (PPC)
- Test inhibition
- Test enhancement
- Experiences with Interferences
- Test enhancement
- Endotoxin Masking
- Planning and implementation of hold-time studies
- Interpretation of hold-time studies
- Mechanistic principles of demasking
- Development of demasking protocols
- Implementation of demasking protocols
- Practical demasking procedure
- Preparation of reagents for demasking
- Application of demasking protocols
- Microbiological Control
- The new science of immunological control
Simulation of contamination in various sample matrices
- Preparation of samples affected by: Test interference, Sample interference
Analysis of interference in affected samples
- Application of different detection systems
- Limulus Amebocyte Lysate assay
- Recombinant Factor C assay
Sample treatment for demasking
- Screening for demasking protocol
- Optimization of demasking protocol
- Evaluation of demasking protocol
Interpretation and Comparison of Results
- Differentiation between test and sample interference
- Effects of different detection systems
- Demasking of endotoxin
Closing Remarks (ECA Academy)