2018 PDA Endotoxins Workshop

The use of recombinant factor C (rFC) for endotoxin detection has been available for many years as an alternate to LAL-based testing.  However, rFC has not gained a proportional use to date, although new trends suggest this might be changing. There are at least two sources of rFC available, and recent publications that suggest it performs similarly on actual biotech samples to LAL. In some cases, there are advantages afforded rFC. The purpose of this section is to review the background regarding the rFC invention and the data from industry comparing it to LAL, as well as the overall burden of changeover. Other factors, such as sustainability of using a natural animal source for LAL will be discussed, as well as future thinking about where our industry might be headed.

2:00 p.m. – 2:30 p.m.
rFC – From Genetic Engineering to Biomedical/Industrial Applications in Endotoxin Detection
Lynne JL Ding, PhD, Professor, National University of Singapore

2:30 p.m. – 3:00 p.m.
rFC – The Road Less Traveled:  An Implementation Update and Shared Learning
Jay Bolden, Senior Consultant Quality-Biologist, Eli Lilly and Company

3:00 p.m. – 3:30 p.m.
Questions and Answers/Discussion

When present in parenteral products, microbial pyrogenic substances (also referred to as Pathogen Associated Molecular Pattern” [PAMP]) can cause life threatening fevers. The human response to PAMPs is regulated by monocytes. The activation starts with PAMP binding to toll-like receptors (e.g. TLR2, TLR4, TLR7, TLR8, TLR9) and involves a cascade of intracellular reactions resulting in a release of pro-inflammatory cytokines. The traditional methods for pyrogen detection either require animal use for the Rabbit Pyrogen Test (RPT), or are limited to the detection of endotoxin (LAL). This session will describe considerations for implementation of the MAT in the pharmaceutical industry.

4:00 p.m. – 4:30 p.m.
MAT: The Stony Way to the Stars
Peter Bruegger, Head Bioassay & Microbiology, Lonza AG

4:30 p.m. – 5:00 p.m.
MAT: How to Conduct a Risk Analysis and Develop a Test Protocol
Ned Mozier, PhD, Senior Director, Analytical R&D, Pfizer Inc.

5:00 p.m. – 5:30 p.m.
Questions and Answers/Discussion

Endotoxin removal by depyrogenation is key to successful parenteral manufacturing. In recent years, USP has dedicated 4 chapters to depyrogenation. This session will discuss general depyrogenation, dry heat depyrogenation, depyrogenation by filtration and endotoxin indicators. A formal risk analysis will be done for the removal of endotoxin in a basic parental process.

8:30 a.m. – 8:50 a.m.
Industry Perspective on Depyrogenation
IJames E Akers, PhD, President, Akers Kennedy & Associates

8:50 a.m. – 9:10 a.m.
Depyrogenation of Primary Packaging - Pharmaceutical Washing Process Validations.
Antje Hermelink, Sr Manager Laboratory GER, Global SME, West Pharmaceutical Services

9:10 a.m. – 9:30 a.m.
Case Study: Dry-Heat Depyrogenation of Glass Articles
Veronika S. Wills, Associate Manager, Technical Services, Associates of Cape Cod, Inc.

9:30 a.m. – 10:00 a.m.
Questions and Answers/Discussion

There are many factors that can stimulate the innate immune systems. Historically, analytical testing has focused on fever and the responses of the innate immune system, which are immediate and measurable responses. However, with the advent of biologics, the adaptive immune system responses have become more and more important. The innate immune system has been found to trigger adaptive immune responses. The growing knowledge of the interface between innate and adaptive immunity needs to be explored to adjust current microbial control concepts. Microbial impurities of parenterals can induce significant immune responses and negative effects on patient safety. In contrast, for vaccines, the presence of “microbial impurities” also may have beneficial effects. Adding defined amounts of stimulating components, such as “detoxified” LPS to the product has shown to improve efficacy. The mechanisms behind this practice are still not fully understood and further research is needed. In this session, innate immune response-modulating impurities are explained and strategies for their control are discussed.

10:30 a.m. – 11:00 a.m.
In Vitro Tools to Control Product Quality Attributes Related Risk Assessments for Ocular Biologic Drugs
Quin (Joe) Zhou, PhD, Associate Director, Allergan

11:00 a.m. – 11:30 a.m.
Endotoxin in the Adaptive Immune Response
Kevin Willliams, Senior Scientist, bioMérieux

11:30 a.m. – 12:00 p.m.
Questions and Answers/Discussion

1:30 p.m. – 3:00 p.m.
Bacterial Endotoxin Testing and Data Integrity

• Session 1: 1:30 p.m. – 2:00 p.m.
• Session 2: 2:00 p.m. – 2:30 p.m.
• Session 3: 2:30 p.m. – 3:00 p.m.

Attendees will participate in all breakouts, for 30-minutes each.

Data integrity continues to be an area of concern among industry and regulatory agencies. The five ALCOA principles encompass all quality data, including data generated using bacterial endotoxin test. In these breakout sessions, data integrity case studies and risk assessments using gel clot, kinetic, and automated endotoxin testing platforms will be presented. Additionally, attendees will participate in small group discussions to better understand data integrity and bacterial endotoxin testing.

Members of PDA’s LER Task Force will present an overview of the three main topics from the LER technical report: 1) the underlying mechanisms and contributing factors of LER, 2) the potential clinical impact of the LER phenomenon, and 3) guidelines for developing LER hold-time study design and LER mitigation strategies. Following the technical report overview will be a discussion of a practical example of addressing LER in existing commercial products.

3:30 p.m. – 3:45 p.m.
LER Technical Report Overview
Friedrich von Wintzingerode, PhD, Senior Manager Global Analytical Science & Technology Microbiology, Roche-Genentech

3:45 p.m. – 4:05 p.m.
Proposed Mechanism for LER
Jack Levin, MD, Professor of Laboratory Medicine and Medicine, University of California School of Medicine, San Francisco

4:05 p.m. – 4:15 p.m.
Q&A with Jack Levin, MD

4:15 p.m. – 4:35 p.m.
Potential Safety Impact of LER and LER Mitigation Strategies
Ned Mozier, PhD, Senior Director, Analytical R&D, Pfizer Inc.

4:35 p.m. – 4:45 p.m.
Q&A with Ned Mozier, PhD

Members of PDA’s LER Task Force will present an overview of the three main topics from the LER technical report: 1) the underlying mechanisms and contributing factors of LER, 2) the potential clinical impact of the LER phenomenon, and 3) guidelines for developing LER hold-time study design and LER mitigation strategies. Following the technical report overview will be a discussion of a practical example of addressing LER in existing commercial products.

5:00 p.m. – 5:20 p.m.
Industry Perspective on LER
Anders Thorn, Principal Scientist, Microbiologist, Novo Nordisk A/S

5:20 p.m. – 5:30 p.m.
Q&A with Anders Thorn

5:30 p.m. – 5:50 p.m.
Case Study: Low Endotoxin Recovery for Existing Commercial Products
Mark J. Kapeckas, Biologics Corporate Quality Program Manager, Sanofi Biologics

5:50 p.m. – 6:00 p.m.
Q&A with Mark J. Kapeckas