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 The 2015 Bacterial Endotoxin Summit (BES) is the 10th annual meeting in a series devoted to the dissemination of non-commercial, scientific information on various topics of interest to scientists interested in the Bacterial Endotoxins Test.

 

The theme for this year's meeting is "Low Lipopolysaccharide and Endotoxin Recovery."   The speakers include a wide range of industry, vendor and regulatory participants. Representatives from FDA, USP, EP, JP and key industry opinion leaders are scheduled to participate. 

 

Questions?  Please contact us by Email at  conference@microbiologynetwork.com or by phone at +1 585-594-3336.

Conference Location:   Iselin, NJ

Venue:

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Renaissance Woodbridge Hotel
515 US Hwy 1 S
Iselin, NJ 08830   USA

Book room at the group rate of $139/night for the Bacterial Endotoxin Summit


Rooms at the group rate are limited.  Early room reservations are recommended.

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Nearest airport:  Newark Liberty International Airport (EWR)

We suggest waiting to make airline reservations until you receive notification that the minimum number of registrants has been met and the conference is confirmed.

Agenda for the 2015 PMF Bacterial Endotoxin Summit -

Low Lipopolysaccharide and Endotoxin Recovery 

September 14, 2015

Time

Speaker

7:30-8:00 am Registration and continental breakfast

8:00-8:30 am

Welcome
Karen McCullough, MMI

8:30-9:45 am

Low endotoxin recovery and how to address it
Michael Dawson, PhD

9:45-10:00 am

Break

10:00-11:15 am

Visualized LPS morphology changes using high speed atomic force microscopy
Masakazu Tsuchiya, PhD

11:15-12:30 pm

Understanding the principles of endotoxin masking and de-masking
Johannes Reich, PhD and Holger Gallert

12:30-1:30 pm

LUNCH

1:30-2:45 pm

Holistic Approach to control: Impacts of low endotoxin recovery
Joseph Chen, PhD and Friedrich von Wintzingerode, PhD

2:45-3:00 pm

Break

3:00-4:15 pm

LPS and LER in the Context of Preventing Microbial Contamination Events in Drug Manufacturing
Kevin Williams

4:15-5:00 pm

Panel Discussion

  • Dennis Guilfoyle, PhD
  • James Cooper PharmD
  • Kevin Williams
  • Mick Dawson, PhD
  • Karen McCullough
  • Johannes Reich, PhD
  • Joseph Chen, PhD

5:00-6:30 pm

Reception

 

 

September 15, 2015

Time

Speaker

8:00-8:30 am Registration and continental breakfast

8:30-9:45 am

Biological Product Matrices and LLR vs LER: What is clinically relevant?
Cheryl Platco

9:45-10:00 am

Break

10:00-11:15 am

Low LPS and Endotoxin Recovery: Keeping an eye on the big picture
Jay Bolden

11:15-12:00 pm

Panel Discussion - LER . . . Just When you Thought You Were Safe
Jeanne Mateffy, Matthew Nelson, Jack Negron-Nieves

12:30-1:30 pm

Lunch

1:30-2:45 pm

Ten Things That the FDA Believes You Should Know About Low Endotoxin Recovery (LER)
Marla Stevens-Riley, Ph.D., FDA

2:45-3:45 pm

Round Table

  • Jay Bolden
  • Joseph Chen, PhD
  • James Cooper, PharmD
  • Mick Dawson, PhD
  • Dennis Guilfoyle, PhD
  • Jeanne Mateffy
  • Karen McCullough
  • Matt Nelson
  • Cheryl Platco
  • Marla Stevens-Riley, PhD
  • Masakazu Tsuchiya, PhD
  • Kevin Williams
3:45 - 4:00 pm

Concluding Comments
Karen McCullough

4:00 pm

Conference Ends

 

Abstracts for the 2015 PMF Bacterial Endotoxin Summit -

Low Lipopolysaccharide and Endotoxin Recovery


Low LPS and Endotoxin Recovery: Keeping an Eye on the Big Picture
Jay Bolden, Eli Lilly

The low LPS issue is approaching 3 years of industry and regulatory activity. Key learning during this time appears to have limited the scope of the issue to the use of LPS as an analyte in hold studies for chelating matrices common to biologic biotech products. Study data and experiences are presented here to share knowledge, explore and challenge fundamental LPS and endotoxin hold study design and associated methodologies. 

 


Holistic Approach to control: Impacts of Low Endotoxin Recovery
Joseph Chen, PhD, and Friedrich von Wintzingerode, PhD
  
 Since Chen first reported the Low Endotoxins Recovery (LER) phenomenon in April 2013, more than 50% of biologic products including monoclonal antibodies and therapeutic proteins in recent FDA BLA submissions were impacted by LER and required by the health authority to implement extensive endotoxins control including addition of the rabbit pyrogen test as an interim product release measure, tightening specifications and risk assessment of manufacturing processes where endotoxin values cannot be accurately determined by LAL assays. Despite several firms reported that LER can be mitigated by Naturally Occurring Endotoxins (NOE), our data show NOE can only delay the onset of LER in biologic product but not a full mitigation. Alternative bacterial endotoxins and pyrogen test methods such as Recombinant Factor C, EndoLISA® and Monocyte Activation Test as well as sample treatments using divalent cations and Endo-RS® demasking kit were evaluated as analytical solutions to LER for Roche/Genentech biologics and none of the described approaches can effectively overcome the LER phenomenon. By collaboration with an academic institute, Roche/Genentech has developed an analytical method to achieve consistent recovery of the spiked CSE in the LER impacted product. We will present a holistic approach to address the LER impacts and describe the analytical solution in details.
.


Low endotoxin recovery and how to address it 
Michael Dawson, PhD, 

 


LER...Just When You Thought You Were Safe 
Jeanne Mateffy, Matthew Nelson, Jack Negron-Nieves

In 2013 Amgen initiated bacterial endotoxin hold time studies for legacy products after the 2012 publication of the FDA Guidance for Industry, Pyrogen and Endotoxin Testing: Questions and Answers document. Initial hold-time study results from the standard Amgen monoclonal antibody platform drug products showed excellent recovery of the spiked control standard endotoxin (CSE) over a 14 day hold period. After attending multiple meetings and reading journal publications where other companies spoke about Low Endotoxin Recover (LER), though sympathetic, Amgen felt relatively safe that we had dodged the LER bullet.

In early 2014 Amgen performed an LER hold time study on a new drug product that is different than our normal MAB platform products and we were thrown headfirst into the LER controversy. At first we thought it was our technique, but we soon learned that the final drug product, drug substance, and intravenous reconstitution fluid all showed the LER effect within minutes of the spike hold time studies. The studies were repeated successfully with a naturally occurring endotoxin (NOE), showing very stable recovery but, regulatory authorities did not accept that data, leading to the performance of rabbit pyrogen studies. The rabbit pyrogen studies were performed with CSE spiked samples and unexpectedly showed a positive reaction, which lead to the implementation of rabbit pyrogen testing for release of the drug product and the IV reconstitution fluid.

In this Round Table Amgen will discuss the studies that were performed as part of the LER investigation, including some interesting nuances. The hope is that other companies will share their LER stories and as an industry we can come up with an assay that is not affected by low endotoxin recovery.  


Biological Product Matrices and LLR vs LER: What is clinically relevant?
Cheryl Platco

The usefulness of recovering purified LPS from biological matrices is being challenged since true endotoxin is the analyte which is detected. There are a variety of mechanisms which can interfere with detection of endotoxin. There are also mechanisms and product matrices which actually destroy or otherwise alter the endotoxin reactivity, rendering the analyte non reactive in the LAL, MAT test and in vivo RPT. The author will examine the clinically relevant detection of true endotoxin versus the clinically irrelevant detection of artificial purified LPS in a variety of biological matrices.
 

Understanding the Principles of Endotoxin Masking and Demasking
Johannes Reich, Ph.D. and Holger Grallert

The detectability of endotoxin contaminations in parenteral drugs is a widely debated, hot topic in the pharmaceutical industry. In recent presentations and publications, the issue of endotoxin masking (Low Endotoxin Recovery) has been discussed. Several experiments have shown that endotoxins can be masked over time, resulting in inadequate activation of LAL-based test methods.

The driving forces of endotoxin masking have been identified as interplay of chelating agents and self-assembly properties of amphiphilic molecules. However, depending on the origin and preparation of endotoxin, variances in masking were observed. Here, we compared different endotoxins with regard to their capabilities in being masked. For example, endotoxins from E. coli O113 were quickly masked, whereas endotoxins from B. cepaci were hardly affected by masking under the same experimental conditions.

Understanding the mechanism of Endotoxin masking enabled us to develop dedicated sample preparation protocols for demasking. Thereby, an essential prerequisite for demasking is the re-arrangement of a detectable Endotoxin aggregation state. Here, we demonstrate the applicability of these protocols and give an insight into the process of demasking.


Visualized LPS morphology changes using high speed atomic force microscopy
Masakazu Tsuchiya, PhD, Charles River Laboratories

 To elucidate the mechanism of the Low Endotoxin Recovery (LER), morphological changes of LPS under the LER condition were observed by using high speed atomic force microscopy (HSAFM).  HSAFM, a recently developed high speed and high resolution scanning probe microscopy by using intermittent contact mode (tapping mode), allowed us to observe real time change of LPS aggregation under a LER condition.  LPS purified from E. coli O55:B5 in water showed a few large (ca. 200 nm long, 20-30 nm wide, 10 nm high), some middle (ca. 60-80 nm long, 10-20 nm wide, 8 nm high), and plenty of small (ca. 20-30 nm long, 10-20 nm wide, 8 nm high) aggregates.  The sizes of aggregates were changed to plenty of small size aggregates (10-20 nm long, 5-7 nm wide, 8 nm high) in 10 mM sodium citrate and 0.05% Polysorbate 20.  Shaving of LPS aggregations was observed by addition of citrate and Polysorbate to LPS in water.  The LPS aggregates seemed to be very soft and fragile in citrate and Polysorbate because scanning by the cantilever on the HSAFM seemed to enhance the shaving of the LPS aggregation.  Since the force on the surface from the cantilever was less than 100 pN, citrate and Polysorbate made the LPS aggregation fragile.  Considering these observations, the mechanism of LER can be as below;

  1. A chelating agent remove magnesium and calcium from LPS aggregations, and this makes LPS aggregations fragile.
  2. A detergent is shaving LPS molecules from the fragile LPS aggregations.
  3. Assuming that there is a threshold of the LPS aggregation size showing high biological activity, the biological activity of the LPS aggregations is significantly reduced when most of the LPS aggregations become smaller than the threshold size. 

LPS and LER in the Context of Preventing Microbial Contamination Events in Drug Manufacturing
Kevin Williams

LPS and LER in the Context of Preventing Microbial Contamination Events in Drug Manufacturing
• Definitions of adulteration
• The use of LPS as a quality indicator
• Surveying some historical contamination events
• Where LER may or may not rise to the level of adulteration
• Do monomers matter?
• “Right-sizing” LPS for detection
 


Ten Things That the FDA Believes You Should Know About Low Endotoxin Recovery (LER)
Marla Stevens-Riley, Ph.D., FDA

Biologic license application drug products intended for injection are required to be tested for pyrogenic substances prior to release per 21 CFR 610.13(b). Many sponsors choose to implement the use of the USP <85> Bacterial Endotoxins Test per 21 CFR 610.9 to fulfill this requirement. However, in recent years questions have arisen regarding the ability of this test to accurately detect the presence of endotoxins in some formulations of biologic license finished drug products due to low endotoxin recovery in spiked drug product samples.  This phenomenon, called Low Endotoxin Recovery (LER), is a concern to the FDA because it implies that endotoxin contamination that could occur during manufacturing might not be detected in the product using the currently available test methods. This talk will outline some points that FDA would like industry to consider regarding endotoxin testing and biologic license drug products.

 

Speakers for the 2015 PMF Bacterial Endotoxin Summit -

Low Lipopolysaccharide and Endotoxin Recovery

 


Jay Bolden
Eli Lilly
Mr. Jay Bolden is an Associate Senior Consultant Biologist in the Eli Lilly and Company Global Quality Laboratories. He is an internal endotoxin subject matter expert and leads a team with global QC oversight for developing, validating, transferring and troubleshooting microbiology, endotoxin, bioassay, QPCR and ELISA methodologies, supporting over 15 sites globally and additional external manufacturing and laboratory partners. 
 
Jay holds a B.S. in Biology and an Environmental Studies certificate from Indiana University, and has 13 years of industry experience in development, process and laboratory microbiology, and microbiology laboratory leadership.  
 

Joseph Chen, PhD
Director and Head of Microbiology Method Management & Technology
Global Biologics Quality Control
Genentech Inc.
 
 

Dr. Chen is the head of Microbiology in Global Biologics Quality Control at Roche and Genentech responsible for both Microbiology and Molecular Biology functions. This includes operational and technical QC method development, validation and maintenance for 10 biologics production sites and for Genentech’s commercial biologics products and IMP products manufactured at CMOs.

Previous to this role he was the head of Quality Control in Genentech Oceanside DS Manufacturing site from 2006-2008. Prior to Genentech, he was the Principle Scientist and Associate Director of Microbiology at Johnson & Johnson, where his responsibilities focused on Research and Commercial Product Safety and Strategic Planning.

Dr. Chen did his graduate study at UCSF with Dr. Jack Levin, the founder of Bacterial Endotoxins LAL assay and holds a PhD in Microbiology and Immunology from University of Rochester in New York. He has 20+ years of business and biopharmaceutical experience, including R&D, quality control, product safety compliance, regulatory CMC and risk management.  


James F. Cooper, PhD
Endotoxin Consultant
 
James F Cooper, PharmD, was Professor of Pharmacy at the Medical University of South Carolina before founding Endosafe in 1987.   This year he received the James P. Agalloco Award for teaching excellence at PDA. His thesis work under Dr. Jack Levin at The Johns Hopkins University in 1971 led to application of the bacterial endotoxin test (BET) for parenteral products. His publications span the history of LAL technology. Since retirement in 2001, he continues his consultation, teaching, publishing related to endotoxin issues, BET root-cause investigations and CGMPs for positron-emission tomography drugs. 
 

Michael Dawson, PhD
Associates of Cape Cod
 
 

Dr. Dawson is Director of Regulatory Affairs at Associates of Cape Cod, Inc. (ACC). His responsibilities include world-wide compliance of ACC’s products. He has broad experience in the manufacture and application of endotoxin testing reagents, instrumentation and of software. Dr. Dawson speaks frequently on endotoxin related issues. He has directed and participated in many courses on the bacterial endotoxins test and the regulatory aspects of its application in health care industries. He has authored many publications, edits the ACC newsletter, LAL Update, and has served on expert panels and committees.

Dr. Dawson received his Bachelors degree from the University of Wales and his doctorate from the University of Southampton, both in the United Kingdom. He is a member of PDA and RAPS and holds Regulatory Affairs Certification (RAC) for the United States.

 

Dennis Guilfoyle, Ph.D. 
Sr. Director, Microbiology & Analytical Regulatory Compliance, Johnson & Johnson

Retired from FDA after 41 years in Seprtember of 2014, Dr. Guilfoyle is now with Johnson & Johnson.
 
Dr. Guilfoyle was classified as a FDA International expert in pharmaceutical microbiology.
He serves as an instructor at both national and regional FDA training courses for both pharmaceutical and biotechnology issues. He performs pre-approval inspections for NDAs\(A)NDAs, and has assisted on over 100 team inspections that include Team Biologic and Medical Device assignments. He is also a member of the FDA Process Analytical Technology Review and inspectional cadres for microbiological applications. Dr. Guilfoyle has testified and consulted as a government expert for the U.S. Attorney's Office on several FDA court cases (civil and criminal). He has co-authored 28 scientific publications and co-wrote/reviewed several FDA field guidelines for microbiological inspections as well as some industry technical baseline documents.
 
Dr. Guilfoyle is active in PDA and has been an active member of the USP Microbiology Committee of Experts for over 10 years.   In addition to Dr. Guilfoyle's insdustry experience he has been an adjunct profession for St. John's University for many years.
 

Karen McCullough
MMI
 
Karen Zink McCullough is principal consultant at MMI Associates, a consulting firm specializing in Endotoxin and Microbiology Testing, GMP Auditing and Compliance, Laboratory Auditing and Compliance and Quality Assurance including nonconformance investigation and CAPA.  Ms McCullough has over 35 years of experience in the Pharmaceutical Industry, and has worked in aseptic manufacturing, oral dosage, and diagnostics sectors. She is a highly respected speaker and trainer both nationally and globally.  Karen's current interests are the use of microbiological assays as tools for process control.
 

Ms. McCullough is a charter member and current Chair of the LAL Users' Group.  She is an elected member of the Microbiology General Chapters Expert Committee of the United States Pharmacopeia and serves as a member of the United States delegation to TC2-9, WG02, revision to ISO 14698 (Biocontamination standard).  She recently edited the book, "The Bacterial Endotoxins Test:  A Practical Guide", which provides background and advice on a range of topics related to the compliant performance of the Bacterial Endotoxins Test. 

 

Jeanne Mateffy
Amgen
 
Jeanne is Chief Microbiologist, Director, Quality Sciences group for Amgen Inc.. In her role she supports global microbiology initiatives by setting policy and administering microbial assays, including sterility testing, bacterial endotoxin testing, bioburden, environmental monitoring, and rapid microbial methods. She is the lead for the Microbiology and Endotoxin Technical Excellence Team and business process owner for the Environmental Monitoring Network and a key member of both the Analytics of the Future and Contamination Control work streams.

Jeanne joined Amgen in 2011, after having worked for various companies supporting microbiology issues. Most recently she worked for Merck/Schering-Plough in different roles including Director of Water Compliance, which included developing guidelines, engineering standards, auditing and training in order to remediate over 100 water systems across the company. Additionally she administered the Rapid Microbiology team, was part of the global microbiology group and ran the NJ microbiology laboratories during the S-P Consent Decree.

Jeanne has her MS in Pharmaceutical QA & Regulatory Affairs, from Temple University, a Master’s certificate in Clinical Trial Management from Drexel University, and a BS in Applied Science & Technology with a concentration in Medical Laboratory Sciences from Thomas Edison State College.


Cheryl Platco
Merck
 
Ms. Platco has been employed with Merck & Co. for 29 years. She is currently a research fellow, where she performs microbiological testing for all pharmaceutical and vaccine/protein products from basic research through marketed product. She is experienced with all microbiological compendial testing.

Ms. Platco has been involved with endotoxin testing for 25 years. She has experience with all three of the major U.S. LAL vendors (ENDOSAFE, LONZA and ACC.) She performs testing for pharmaceutical, vaccine, biological, and monoclonal products as well as a variety of raw materials, active ingredients, and culture media.

She currently works at the West Point PA facility in Merck Research Laboratories, Analytical Sciences.

Ms. Platco holds a B.S. in Chemistry/Biology and Medical Technology degree. She also holds an MS in Clinical Chemistry. 
 

Marla Stevens-Riley, Ph.D.
FDA

Dr. Stevens-Riley is currently a senior microbiologist in the Division of Regulations, Guidance, and Standards in the Office of Policy for Pharmaceutical Quality in the Office of Pharmaceutical Quality (OPQ) in CDER. Previously, she was a Team Leader in the Division of Microbiology Assessment in the Office of Pharmaceutical Science in CDER.
She is a CDER liaison to the United States Pharmacopeia (USP) General Chapters-Microbiology Expert Committee, and she is currently one of the co-leaders on the Parenteral Drug Association (PDA) Task Force rewriting the PDA Technical Report 27: Pharmaceutical Package Integrity. She was the lead author on the guidance for industry Submission of Documentation in Applications for Parametric Release of Human and Veterinary Drug Products Terminally Sterilized by Moist Heat Processes and has recently published the paper Parametric Release: A Regulatory Perspective in American Pharmaceutical Review. During her scientific career, she has co-authored over 15 peer-reviewed scientific publications. Dr. Stevens-Riley holds a B.S. in biology from the College of William and Mary and a Ph.D. in Microbiology from the University of Georgia, and she completed post-doctoral training in the Department of Microbiology and Immunology at the University of Texas Southwestern Medical Center in Dallas.

 


Masakazu Tsuchiya, PhD
Charles River Laboratories 

Masakazu “KAZ” Tsuchiya has been a Senior Research Scientist in Endotoxin and Microbial Detection, Charles River since 2007. He has 30 year experience on the detection of endotoxin and other microbial cell wall components in Wako Pure Chemical Industries, Ltd, Osaka, Japan, Duke University Marine Laboratory, Beaufort, NC, Wako Chemicals USA Inc., St. Louis, MO, and Charles River, Charleston, SC. He earned a doctorate of Agriculture and a M. S. in Food Science and Technology from Kyoto University, Japan. His notable achievement is the development of tools for detection of microbial cell wall components, such as an endotoxin-specific LAL reagent, a beta-glucan-specific LAL reagent, and the Silkworm Larvae Plasma (SLP) reagent to detect peptidoglycan and beta-glucan.   


Kevin Williams
Lonza

Expert microbiologist specializing in endotoxin and microbiology analytical development (methods/ instrumentation /software /training), investigations (troubleshooting/ root cause analysis), and cGMP quality solutions and systems (regulatory responses/ white papers/ control strategy/ theory). Developed, published and implemented methods for the determination of specifications for bulk drugs and drug excipients/ raw materials.