Module 9 2024

03/09/2024

Examples of Manufacturing Process CPPs and IPCs

USP Manufacturing Flow Chart

Step Number

Operation

Purpose

CPP

IPC

Thawing of WCB and inoculation in to 125 mL shake flask

USP-1

Vial Thaw and Inoculation

Inoculation cell dilution

Cell viability before dilution

Sequential expansion in shake flasks (125, 250, 1000, 3000 mL)

Culture expansion before inoculation into 50 L bioreactor

Duration, seeding density, rocking speed, angle, CO 2 flow rate

Cell viability before dilution, bio-contamination at harvest, viable cell density, cell viability, bioburden

USP-2

Culture expansion before inoculation into 100 L bioreactor

Inoculation cell dilution, temperature, pH, dissolved O 2 , N 2 flow, media incubation duration

Viable cell density, cell viability, bioburden prior to transfer, biocontamination at harvest

USP-3

Expansion in 50 L bioreactor

Culture expansion before inoculation into 500 L bioreactor

Expansion in 100 L bioreactor

Inoculation cell dilution, temperature, pH, dissolved O 2 , N 2 flow, media incubation duration

Viable cell density, cell viability, bioburden prior to transfer, biocontamination at harvest

USP-4

Culture expansion before inoculation into 2000 L production bioreactor

Expansion in 500 L bioreactor

Inoculation cell dilution, temperature, pH, dissolved O 2 , N 2 flow, media incubation duration

Viable cell density, cell viability, bioburden prior to transfer, biocontamination at harvest

USP-5

Medium storage temperature and duration, dissolved O 2 , seeding density, agitation, duration, temperature, pH, airflow, feed time and quantity

Production in 2000 L bioreactor

USP-6

Bulk production of mAb

Viable cell density, cell viability, glucose profile

Harvest of cell culture supernatant

USP-7

Crude harvest

-

Mycoplasma, bioburden, adventitious viruses

Production of clarified harvest

USP-8

Depth filtration

Filter load, inlet pressure, storage of filter

Endotoxins, bioburden, biocontamination, yield

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Examples of Manufacturing Process CPPs and IPCs

DSP Flow Manufacturing Flow Chart

Step Number

Operation

Purpose

CPP

IPC

Removal of HCPs, DNA, and media components

Temperature, flow rate, storage duration

DSP-1

Affinity chromatography

Endotoxin, bioburden, HCP, HCDNA, purity

Temperature, incubation pH, incubation time

Bioburden

DSP-2

Low pH hold

Low pH virus inactivation

Anion exchange chromatography

Removal of DNA and anionic HCPs

Temperature, flow rate, product load

DSP-3

Bioburden, purity

Cation exchange chromatography

Temperature, flow rate, product load

DSP-4

Removal of cationic HCPs

Bioburden, purity

DSP-5

Virus filtration

Virus reduction

Filter capacity

Bioburden, purity

Adjustment of concentration and buffer exchange

DSP-6

Ultrafiltration / Diafiltration

-

Bioburden, purity, pH, protein concentration

DSP-7

Formulation

Formulation of bulk DS

-

Bioburden, endotoxin, protein concentration

Transfer into DS storage container

Freezing, storage and shipment

DSP-8

Bulk fill

DS Release Tests

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