Bioburden :The Burden

on our
Biological Operations
S.Ballal
Head, Biopharma Bulk Mfg

12/16/09 1
Objective Of This Training
 To understand
 Issues related to Bioburden
 Sources of Bioburden

 To Implement
 Methods & Systems to Control Bioburden

 Ultimate Goal
 To Prevent Loss of Time, Resources and Money

12/16/09 Confidential 2 of 15
Microbiology Primer
 Bioburden:

Population of viable microorganisms on a raw material,

component, a finished product, and/or package.

Measured in CFU (colony forming units) per unit of

product

 Sterile:
 Free from living organisms (Microbes)
 Aseptic
 Removal of Bioburden

12/16/09 Confidential 3 of 15
Where all do we need Bioburden
Control ?
 Raw Material: including water
 Resins
 Membranes
 Intermediates
 Drug Substance/Product – Stability
Samples, Control Samples
 Enzymatic reactions
 Reagents – pH standards, Buffers

12/16/09 Confidential 4 of 15
Why Bioburden Control is
Critical
 Product Degradation /Spoilage
 Safety
 Efficacy
 Appearance
 Material Degradation & Life Cycle
Reduction
 Affect experimental data & Results

12/16/09 Confidential 5 of 15
Solution for Bioburden
Control

Use everything sterile

Commercially and Technically not feasible

and not required too.

12/16/09 Confidential 6 of 15
Sources of Bioburden
 Raw Material
 Processing Equipment
 Environment
 Personnel
 Packing Material

12/16/09 Confidential 7 of 15
 Bioburden Control in R
& D Set up

12/16/09 8
Material Sourcing
 Low Bioburden Products- Expensive
 Material inherently low in bioburden
 Organic Products
 Inorganic salts
 Material inherently high in bioburden
 Aqueous liquids- with / without salts & sugars
 Natural Products – Yeast Extract, Peptone

 Water- Fresh Every Day

12/16/09 Confidential 9 of 15
Raw Material Handling
 Minimize opening and
 Use clean tools for dispensing
 Keep the lid on–reduce moisture ingress
 Refrigerate when instructed/possible
 Avoid low and high bioburden processing together
 Open Critical Material in Controlled air – LFH,
Biosafety hood etc.

12/16/09 Confidential 10 of 15
Process Equipment
 Immediately clean after use
 Use bioburden reduction agents where ever possible –
NaOH, Acids, Hyprochloride
 Use Autoclaved glassware for critical operations
 If in doubt, clean/ sanitize again
 Store either in very dry state or with biostatic liquid- no
not store wet/ moist
 Periodically clean equipment thoroughly for removing soil/
grime from hard to clean areas

12/16/09 Confidential 11 of 15
Environment & Personnel
 Keep Lab area clean
 Use LFH for all critical sample handing- Stability, DS,
PCT DS / DP
 Aliquot when possible to reduce risk
 Following good handing procedures
 Sanitize hands before critical material handling
 Hold containers away from mouth/ brim
 Keep your body and other things away from the
product

12/16/09 Confidential 12 of 15
Controlling Bioburden in
Process
 Filter Buffers and solutions
 Filter Intermediates- for storage
 Filter intermediates before loading on to the
columns
 Filter all critical DS/ DP
 Avoid multiple opening and handling
 Tubing are good source of bioburden – Wet,
difficult to clean
 Use Secondary Packing – Additional Shield

12/16/09 Confidential 13 of 15
Packing Material
 Use Autoclaved Glassware and handling tools

 Always use sterile tips and pipettes

 Falcon tubes are sterile as long as you handle

them that way

12/16/09 Confidential 14 of 15
 When In Doubt –

Use to Stringent
Method for Handling
12/16/09 15