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Challenges in Freeze–Thaw Processing of Bulk Protein Solutions

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Sterile Product Development

Part of the book series: AAPS Advances in the Pharmaceutical Sciences Series ((AAPS,volume 6))

Abstract

Freeze-thawing is a common unit operation during the production of protein-based therapeutics. Bulk protein solutions are often stored in frozen state for extended periods, and thawed to room temperature prior to downstream process operations including lyophilization, add-in formulation ingredients step, and fill and finish processes. However, freezing can induce protein denaturation stresses, such as cold denaturation, ice–liquid interfacial denaturation, and cryoconcentration. Many of these stresses are manifested as unfolding, reversible aggregation, and insoluble particle formation, while some can cause loss of structure and therapeutic function. Numerous studies have been attempted to understand and mitigate protein denaturation during freeze-thawing, and thereby provide guidelines for optimization of formulation and process variables. This chapter presents an overview of freeze-thawing-induced stresses, cryopreservation aspects, and containers, associated with manufacturing of bulk protein solutions.

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Abbreviations

°C:

Degree Celsius

ΔG :

Gibb’s free energy

ANS:

1-Anilino-8-napthalene sulfonate

BSA:

Bovine serum albumin

CMC:

Critical micelle concentration

C p :

Heat capacity

DTPA:

Diethylenetriamine penta-acetic acid

EDTA:

Ethylenediamine tetra-acetic acid

IgG:

Immunoglobulin G

IR:

Infrared

kJ:

Kilojoules

LDH:

Lactate dehydrogenase

LPT:

Last point to thaw

LPTF:

Last point to freeze

LYS:

Lysozyme

mAb:

Monoclonal antibody

mL:

Milliliters

PE:

Polyethylene

PEG:

Poly(ethylene) glycol

PETG:

Poly(ethylene) terephthalate glycol

PFK:

Phosphofructokinase

PP:

Polypropylene

PTFE:

Polytetrafluoroethylene

PVP:

Poly(vinyl) pyrrolidone

rhFXIII:

Recombinant hemophilic factor XIII

rhIFN-γ:

Recombinant human interferon-γ

T c :

Crystallization temperature

T e :

Eutectic temperature

T f :

Equilibrium freezing point

T g :

Glass transition temperature of solids

Tg΄:

Glass transition temperature of frozen solution

T het :

Heterogeneous nucleation temperature

T hom :

Homogeneous nucleation temperature

T m :

Melting temperature

TRE:

Trehalose

Trp:

Tryptophan

T x :

Devitrification temperature

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, The University of Tennessee Health Science Center, 26 South Dunlap Street, Memphis, TN, 38163, USA

    Hari R. Desu Ph.D.

  2. Biotherapeutics Formulation Development, Veterinary Medicine R&D, Pfizer Inc., Kalamazoo, MI, USA

    Sunil T. Narishetty Ph.D.

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Correspondence to Hari R. Desu Ph.D. .

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Editors and Affiliations

  1. Pharmaceutical R&D-BioTx Pharmaceutical, Chesterfield, Missouri, USA

    Parag Kolhe

  2. LifeCell Corporation, Bridgewater, New Jersey, USA

    Mrinal Shah

  3. Drug Product Engineering, Amgen, Thousand Oaks, California, USA

    Nitin Rathore

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Desu, H.R., Narishetty, S.T. (2013). Challenges in Freeze–Thaw Processing of Bulk Protein Solutions. In: Kolhe, P., Shah, M., Rathore, N. (eds) Sterile Product Development. AAPS Advances in the Pharmaceutical Sciences Series, vol 6. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-7978-9_7

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