QA/QC, (c)GMP, SOP, BSL, etc.

1. QA/QC, (c)GMP, SOP, BSL, etc.

2. Quality Control “Quality control is involved in developing systems to ensure products or services are designed and produced to meet or exceed customer requirements and expectations .“

3. Quality Assurance • Covers all activities from design, development, production, installation, servicing and documentation • Includes: • the regulation of the quality of raw materials, assemblies, products and components; • services related to production; and • management, production, and inspection processes

4. PDCA (aka the Shewhart Cycle) • An iterative four-step quality control strategy • Ensure that the product fulfills or exceeds customer expectations • PLAN • establish the objectives and processes necessary to deliver results in accordance with the specifications. • DO  • implement the processes. • CHECK • monitor and evaluate the processes and results against objectives and specifications and report the outcome. • ACT • apply actions to the outcome for necessary improvement.

5. Good Manufacturing Practice • “A set of regulations, codes, and guidelines for the manufacture of drugs (known as medicinal products in Europe), medical devices, diagnostic products, foods products and Active Pharmaceutical Ingredients (APIs).” • cGMP: current GMP

8. Standard Operating Procedures Written descriptions of routine methods (standardize). Referenced or provided in an appendix of the QA project plan. Procedures benefiting from SOPs include: routine measurements chain-of-custody sample handling and shipment routine analytical methods for chemical analyses. Ensure that all persons conducting work are following the same procedures and that the procedures do not change over time. All personnel should be thoroughly familiar with the SOPs before work is initiated. Deviations from SOPs may affect data quality and integrity. If it is necessary to deviate from approved SOPs, these deviations must be documented and approved through an appropriate chain-of-command. Personnel responsible for ensuring the SOPs are adhered to must be identified in the QA Project Plan.

9. Biological Safety Levels(BSLs) “Biosafety Level 1: suitable for work involving well-characterized agents not known to consistently cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environment. The laboratory is not necessarily separated from the general traffic patterns in the building. Work is generally conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is neither required nor generally used. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or a related science. “

10. Microbial Contaminants • Endospores • Quality Degradation • Potential Harm • Endotoxins • Mycoplasmas

11. Bacterial Structures

13. Endospore Formation Process Typically takes hours SURVIVAL! ExtraordinaryResistance!

15. 25 - 40 MYA Bacteria! New Oldest “Living” Endospores - 250 MYA

16. Microbial Contaminants • Endospores • Quality Degradation • Potential Harm • Endotoxins • Harmful effects • Indicate contamination • Mycoplasmas

17. Gram - Cell Wall Composition contains endotoxin *** *** Fig. 3.34

18. Endotoxin Effects

19. Fever (Pyrogenicity):    •   Circulatory system effects: •   Leukopenia followed by leukocytosis: •   Leukopenia: an abnormal reduction in the number of leukocytes •   Leukocytosis: an abnormal increase in the number) of leukocytes •  Increased vascular permeability (vasodilation) •   Decreased peripheral circulation •   Decreased perfusion (blood flow) of blood to major organs •     Effects on blood coagulation: •   (DIC) Disseminated intravascular coagulation: •   Activation of clotting pathway •   Thrombosis: Formation of blood clot (thrombus) in heart or blood vessel    •   Effects on metabolic and liver functions •   Decreased iron availability •   Hypoglycemia: Abnormally low glucose levels •   Cellular death (cytotoxicity) •   Organ necrosis: •   Sum of morphological changes indicative of cell death and caused by the • progressive degradative action of enzymes •   Shock: •   Characterized by failure of the circulatory system to maintain adequate • blood flow to the vital organs •   Symptoms include: Hypotension; Weak pulse; Rapid and shallow breathing; • Low body temperature; CNS (central nervous system) effects (e.g., nausea) •   Death

20. Endotoxin Assay Most commonly used assay: LAL Kinetic Assay - Lymulus (horseshoe crab) amoebocyte lysate (LAL) test Manufactuer’s of LAL:

21. LAL assay background

22. Microbial Contaminants • Endospores • Quality Degradation • Potential Harm • Endotoxins • Harmful effects • Indicate contamination • Mycoplasmas • Harmful effects • Indicate contamination

23. Mycoplasmas

24. Mycoplasma “Silent Infections” “long implicated in a variety of frustrating and expensive cell-culture contaminations.” Lack rigid cell wall not affected by the presence of beta-lactamase antibiotics small in size (0.3 - 0.8 µm) do not produce turbidity very hard to see with light microscope. Presence/growth can result in severe metabolic changes and production of deleterious metabolites affecting valuable data and results. Contamination of cell cultures can result in loss of valuable cell lines, production time and batches of biologicals. Monitoring cell cultures on a routine basis can help identify the source of contaminations as well as prevent future cross-contamination events.

25. Mycoplasma Assays QA/QC Forms

26. General “Sanitary” Monitoring Techniques Petrifilms (3M)