04/08/23 1

Bioburden :The Burden on our

Biological Operations

S.BallalHead, Biopharma Bulk Mfg

04/08/23 Confidential 2 of 15

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

04/08/23 Confidential 3 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

04/08/23 Confidential 4 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

04/08/23 Confidential 5 of 15

Why Bioburden Control is Critical Product Degradation /Spoilage

Safety Efficacy Appearance

Material Degradation & Life Cycle Reduction

Affect experimental data & Results

04/08/23 Confidential 6 of 15

Solution for Bioburden Control

Use everything sterile

Commercially and Technically not feasible and not required too.

04/08/23 Confidential 7 of 15

Sources of Bioburden Raw Material Processing Equipment Environment Personnel Packing Material

04/08/23 8

Bioburden Control in R & D Set up

04/08/23 Confidential 9 of 15

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

04/08/23 Confidential 10 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.

04/08/23 Confidential 11 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

04/08/23 Confidential 12 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

04/08/23 Confidential 13 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

04/08/23 Confidential 14 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

04/08/23 15

When In Doubt –

Use to Stringent Method for Handling