Note: Intro -- Preface -- Contents -- Author Biography -- Part I: Pharma -- Chapter 1: Discovery and Early Development of the Limulus Test -- 1.1 Introduction -- 1.2 Early Observations and Test Development -- 1.3 Official Approval of the Limulus Amebocyte Lysate Test by Federal Agencies -- 1.4 End Points for the Detection of Endotoxin -- 1.5 Conclusion -- References -- Chapter 2: Origins and Evolution of Drug Regulation -- 2.1 Drug Regulations Origin in Tragedy -- 2.2 Biologics Drug Regulation -- 2.2.1 Early Biologics -- 2.2.2 Blood-Borne Concerns: HIV -- 2.2.3 Blood-Borne Concerns: Prions -- 2.3 Biologics Split Between CDER and CBER -- 2.3.1 Categories of Therapeutic Biological Products Transferred to CDER -- 2.3.2 Categories of Therapeutic Biological Products Remaining in CBER -- 2.3.3 Combination Products -- 2.4 Biosimilars -- 2.4.1 Patient Protection and Affordable Care Act -- 2.4.2 Differences in Regulation, Europe Versus U.S -- 2.5 Misadventures in Regulatory and Clinical Pharmacology -- 2.5.1 Compounding Misery -- 2.5.2 Subvisible Particulates -- 2.5.3 Clinical Cytokine Storm -- 2.5.4 Elusively Endotoxin -- References -- Chapter 3: LPS Structure, Function, and Heterogeneity -- 3.1 Introduction -- 3.2 Role of Lipopolysaccharides -- 3.3 Brief History of Lipopolysaccharides -- 3.4 LPS General Structural Architecture -- 3.5 O-Polysaccharides Structures -- 3.5.1 Inter Genera Common or Closely-Related O-chain Structures: The Example of Brucella abortus, Yersinia enterocolitica and Vibrio cholera -- 3.5.2 O-polysaccharide Structure-Function Relationships -- 3.5.3 Core Oligosaccharide Structures -- 3.5.4 Reducing Core Sugars -- 3.5.5 The Sugar Moiety of Bacterial Lipooligosaccharides (LOS) -- 3.6 Core Structure-Function Relationships -- 3.7 Lipid A Structures -- 3.7.1 Number and Length of Fatty Acids -- 3.7.2 Substitution of the Lipid A Phosphate Groups. , 3.7.3 Unusual Lipid A Backbone Structures -- 3.7.3.1 Lipid A with a 2,3-diamino-2,3-dideoxy-D-glucose (DAG) Backbones -- 3.7.3.2 Lipid A with Mixed DAG and GlcN Backbones -- 3.7.3.3 Lipid A with GlcN or DAG Disaccharide Backbones Without Phosphate Groups -- 3.8 Lipid A Recognition Through the TLR-4 Receptor: How Structural Modifications Impact Innate Immunity and Favor Bacterial Virulence and Pathogenicity. Applications to Human Health -- 3.9 Brief Description of the LPS Biosynthetic Pathway -- 3.10 Origin of LPS Heterogeneity -- 3.10.1 Impact of Biofilm Growth Conditions on LPS Structures -- 3.10.2 The Example of the Bordetella Genus LPS: High Structural Diversity and Heterogeneity -- 3.10.3 Lipid A -- 3.10.4 Cores -- 3.10.5 O-chains -- 3.10.6 Bordetella LPS Biological Activities -- 3.11 Other Examples of Heterogeneity and Diversity -- 3.11.1 Example: Salmonella minnesota R595 Re LPS Heterogeneity Shown by Thin Layer Chromatography (TLC), and Matrix Assisted Laser Desorption (MALDI) Mass Spectrometry -- 3.11.2 Example: Vibrio cholerae Lipid A Heterogeneity Shown by MALDI Mass Spectrometry -- 3.12 Conclusions -- References -- Chapter 4: Endotoxin in Microbiological Context -- 4.1 Early and Expanding Microbiological Context -- 4.2 Metazoan Receptors for Microbial PAMPs -- 4.3 Aseptic Processing, Bioburden and Endotoxin Hold Time Studies -- 4.4 Structural Context of LPS in the GNB OM -- 4.4.1 Modeling and Re-modeling of Membranes -- 4.4.2 Constituents of GNB Membranes and Action of Divalent Cations -- 4.4.3 Evolution of GNB OM -- 4.5 The Definition of Endotoxin -- 4.5.1 Biosynthesis of LPS -- 4.5.2 Reciprocal Specificity of the Host Response to LPS -- 4.6 Unique LPS Sugar Moieties by Which GNB Are Defined and Targeted -- 4.6.1 O-Antigen Serotyping -- 4.6.2 Bacteriophage Typing -- 4.6.3 Bacteriophage Transfer of Antibiotic Resistance Genes. , 4.7 LPS Aggregation and Activation of the Immune System -- 4.7.1 The Mueller Study -- 4.7.2 The Takayama Study -- 4.7.3 Refining the Questions -- 4.8 Asymmetry and Proportionality of LPS in GNB OM Structure -- 4.9 Non-LPS OM Constituents Are Dominated by Porins -- 4.9.1 Predominance of Porins -- 4.9.2 Porin Structure and Specificity Contrast LPS -- 4.9.3 Porin Host Reactivity -- 4.9.4 Porin Association with LPS -- 4.9.5 Porin Occurrence in Human Cells and Disease -- References -- Chapter 5: Endotoxin and Microbiological Control -- 5.1 Microbiological Control Mechanisms of Action -- 5.1.1 LPS contribution to OM Impermeability -- 5.1.2 Antibiotic/Antimicrobial Peptides -- 5.1.3 OMV Formation -- 5.1.4 Miscellaneous Perturbation -- 5.1.5 Disinfection -- 5.2 Glass, Increasing Control Requirements -- 5.3 Depyrogenation -- 5.3.1 Preclusion -- 5.3.2 Removal -- 5.3.2.1 Filtration -- 5.3.2.2 Two-Phase Micellar Solution -- 5.3.2.3 Hydrophobic Adsorption -- 5.3.2.4 Chromatography -- 5.3.2.5 Misc. Binding -- 5.3.3 Inactivation -- 5.3.3.1 Dry Heat -- 5.3.3.2 Chemical Methods -- 5.3.3.3 Plasma Gas -- 5.3.3.4 Radiation -- 5.4 Example Projects -- 5.4.1 Medical Devices -- 5.4.2 Nanoparticles -- 5.4.3 Purified Water -- 5.4.4 Biofilm Disruption -- References -- Chapter 6: Practical Limulus-Based Testing -- 6.1 Introduction: Why the Focus on Endotoxin? -- 6.2 Part I. A Well-Supported Test -- 6.2.1 Overview of the cGMP Environment -- 6.2.2 Lab Variability -- 6.2.3 Test Basics: Materials and Methods -- 6.2.4 Sampling and Storage -- 6.2.5 Endotoxin Standardization -- 6.2.6 Out with Old Guidance, in with New -- 6.2.7 Installing a New Test System in a cGMP Environment -- 6.2.7.1 What Steps Are Needed to Setup and Qualify a New Test System? -- 6.2.7.2 Test Type Overview -- 6.3 Part II. A Well-Developed Test -- 6.3.1 Developing a Drug Test. , 6.3.1.1 Calculations Based on Drug Dosage, Method Development Testing -- 6.3.2 Excipient Specifications to Support a Control Strategy -- 6.3.2.1 Calculations to Determine API, Excipient and Raw Material Specifications -- 6.3.2.2 Validation/Verification -- 6.3.2.3 Routine Test Protocol -- 6.4 Part III. A Well-Performed Test -- 6.4.1 The Analyst Test Interface -- 6.4.1.1 Software Template Setup -- 6.4.1.2 Preparation of the CSE/RSE and the Resulting Dilution Series -- 6.4.1.3 Preparation of the Sample -- 6.4.1.4 Addition of Spikes to the Plate -- 6.4.1.5 Addition of Solutions to the Plate -- 6.4.1.6 Hot and Cold Wells -- 6.4.1.7 Pre-incubation of the Plate -- 6.4.1.8 Addition of LAL/rFC to the Plate Containing Standards and Sample Dilutions -- 6.4.1.9 Result Calculation -- 6.4.1.10 Analyst Documentation of Test Performance -- 6.4.1.11 Post-test Activities (pH, Preservation of Sample for Retest or Resample) -- 6.4.1.12 Troubleshooting -- 6.4.1.13 Retest: Resample Performance -- 6.4.1.14 Beta Glucans -- References -- Chapter 7: Emerging Immune Context -- 7.1 Introduction -- 7.2 Emerging Immune Context -- 7.2.1 Immune Reactions Including Fever Are Common in Biologics Administration -- 7.2.2 IIRMI as a New Paradigm -- 7.2.3 Best Proof of Concept -- 7.3 Innate Activation of Adaptive Immunity -- 7.3.1 Microbiology Meets Immunology -- 7.3.2 Receptors and Markers Overview -- 7.3.2.1 CD Markers -- 7.3.2.2 Clonal Versus Non-clonal Receptors -- 7.3.2.3 Fc Receptors -- 7.3.3 PAMP Activation of Adaptive Immunity -- 7.3.3.1 The Second Signal -- 7.3.3.2 Licensing the Second Signal -- 7.4 A Simple Immunological Model for MCC -- 7.4.1 Explanatory Power of a Simple Model -- 7.4.2 Co- Receptor Modulation Is Inherent in Disease Causation as Well as the Mode of Action of Biologics Molecules -- 7.4.3 Action of Adjuvant Usage in Vaccinology. , 7.4.4 Suppression of Host Immunity by Pathogens Via Modulation of Receptors -- 7.5 Specific Ways Adaptive Immune Context Differs from Microbiological Context -- 7.5.1 Synergistic Contaminants -- 7.5.2 Relevant Levels Are Not Necessarily Pyrogenic Levels -- 7.5.3 Time Is a Factor -- 7.5.4 What Is the Meaning? -- 7.6 Milestone Publications in Bridging Innate and Adaptive Immunity -- Appendix I -- References -- Chapter 8: The Biologics Revolution and Endotoxin Test Concerns -- 8.1 Part I Biologics Overview -- 8.1.1 Introduction -- 8.1.2 Differences in Biologics Versus LVPs and SMDs -- 8.1.2.1 Large, Complex and Produced in Living Organisms -- 8.1.2.2 Produced via Recombinant Methods -- 8.1.2.3 Adverse Responses -- 8.1.2.4 The Process Is the Product -- 8.1.2.5 Based Upon Immune Modulation -- 8.1.2.6 Biologics Drug Classes -- 8.1.2.7 Clinical, Regulatory, Legal and Manufacturing Cost Differences -- 8.1.2.8 Require Additional Microbiological Control: Expression System Removal -- Microbial Mimetics Mimetics are not actual microbiological byproducts or cell constituents but rather process constituents that may be immune activating in the body, as surveilled by the immune system, and treated as if it were of a microbial na -- Host Cell Protein (HCP) Impurity Determination HCP removal is applicable to biologics as produced from living cells. As these cells grow and produce therapeutic proteins, they also produce other substances (simply by being alive, reproducing, etc. -- Nucleic Acid Removal According to Wang et al. [53] biologics production has some unique aspects of concern in the removal of nucleic acids, particularly residual DNA:. , Beta Glucans Fungal cell wall constituents, especially beta glucans, have come to be considered by many to be innate immune response modulating impurities (IIRMI) and should therefore be removed or precluded from biologics manufacturing processe.

Note: Front cover -- Preface -- Contents -- Contributors -- Chapter 1. Historical and Emerging Themes -- Chapter 2. Endotoxin Relevance and Control Overview -- Chapter 3. Fever and the Host Response -- Chapter 4. Endotoxin Structure, Function, and Activity -- Chapter 5. Descent of Limulus: Arthropoda and the New Biology -- Chapter 6. Microbial Biodiversity and Lipopolysaccharide Heterogeneity: from Static to Dynamic Models -- Chapter 7. Nonendotoxin Microbial Pyrogens: Lesser Endotoxins and Superantigens -- Chapter 8. Risk Assessment in Parenteral Manufacture -- Chapter 9. Endotoxin as a Standard -- Chapter 10. Mechanism, and Application -- Chapter 11. Development, Validation, and Regulation -- Chapter 12. The Pyrogen Test -- Chapter 13. Pyrogenicity Case Studies -- Chapter 14. Developing Specifications for Active Pharmaceutical Ingredients, Excipients, Raw Materials, Sterile Pharmacy Compounds, and Nutritional Supplements -- Chapter 15. Depyrogenation Validation, Pyroburden, and Endotoxin Removal -- Chapter 16. Automation, Process Analytical Technology, and Prospective Testing -- Chapter 17. Limulus Amebocyte Lysate Testing of Medical Devices -- Chapter 18. Receptors, Mediators, and Mechanisms Involved in Bacterial Sepsis and Septic Shock -- Index.