Publication: Development of a high concentration cell bank production method
Abstract
Biyofarmasötik sektörü, geniş ürün yelpazesiyle genel ilaç pazarının önemli ve büyüyen bir bölümünü temsil etmektedir. Endüstriyel ölçekte üretilebilen memeli hücre kültürü, rekombinant proteinler için en yaygın üretim sistemi haline gelmiştir. Tipik bir memeli hücre kültürü, ilgili hücre bankasından bir vialin çözülmesi, biyokütle miktarının üretim için yeterli düzeye getirilmesi ve üretim aşamalarını içerir. Tohum dizisi, üretim biyoreaktörünü aşılamak ve protein üretimini optimize edilmiş bir şekilde başlatmak için yeterli biyokütle üreten biyoprosesin kritik bir parçasıdır. Geleneksel bir tohum dizisinde bu, hücrelerin çalışan bir hücre bankası vialinden, çalkalayıcı şişeler, sallama hareketli biyoreaktörler ve karıştırmalı tank biyoreaktörleri dahil giderek daha büyük ölçekli yetiştirme sistemlerinden geçirilmesiyle elde edilir. Bu ara-kültür adımlarını azaltmak ve bir üretim biyoreaktörünün doğrudan kriyoviallerden veya kriyotorbalardan alınan hücrelerle doğrudan aşılanmasını sağlayacak düzeyde bir yüksek konsantrasyonlu hücre bankası oluşturmak için çalışmalar halen devam etmektedir. Memeli hücreler için ortalama hücre bankası konsantrasyonu 10-15x106 hücre/ mL'dir. Bu tez çalışmasında, 200x106 hücre/ mL'ye kadar farklı konsantrasyonlarda hücre bankaları oluşturularak doğrudan üretime aşılanmıştır ve kültürlerin büyüme profilleri ve üretilen monoklonal antikor miktarı belirlenmiştir. Bu çalışmanın temel amacı, yüksek konsantrasyonlu hücre bankaları kullanarak zaman, iş gücü, maliyet ve operasyonel riskleri azaltmaktır.
With its diverse product range, the biopharmaceutical sector represents a significant and growing segment of the overall pharmaceutical market. Mammalian cell culture that can be produced on an industrial scale has become the dominant production system for recombinant proteins. A typical mammalian cell culture includes the steps of thawing a vial from the relevant cell bank, bringing the amount of biomass to a sufficient size for production, and production steps. The seed train is a critical part of the bioprocess, producing enough biomass to inoculate the production bioreactor and start protein production in an optimized manner. In a conventional seed train, this is achieved by passing cells through a working cell bank flask through increasingly larger cultivation systems including shaker flasks, rocking motion bioreactors, and stirred tank bioreactors. Efforts are being made to create a high-concentration cell bank to avoid these intermediate culture steps and to allow direct inoculation of a production bioreactor with cells from cryovials or cryobags. The common cell banking concentration for mammalian cells is 10-15x106 cells/ mL. In this study, cell banks with different concentrations up to 200x106 cells/ mL were created and these cell banks were inoculated directly into production to test the growth profiles of the cells and the amount of monoclonal antibodies produced. The ultimate goal of this study was to reduce time, labor, cost and operational risks by using high concentration cell banks.
With its diverse product range, the biopharmaceutical sector represents a significant and growing segment of the overall pharmaceutical market. Mammalian cell culture that can be produced on an industrial scale has become the dominant production system for recombinant proteins. A typical mammalian cell culture includes the steps of thawing a vial from the relevant cell bank, bringing the amount of biomass to a sufficient size for production, and production steps. The seed train is a critical part of the bioprocess, producing enough biomass to inoculate the production bioreactor and start protein production in an optimized manner. In a conventional seed train, this is achieved by passing cells through a working cell bank flask through increasingly larger cultivation systems including shaker flasks, rocking motion bioreactors, and stirred tank bioreactors. Efforts are being made to create a high-concentration cell bank to avoid these intermediate culture steps and to allow direct inoculation of a production bioreactor with cells from cryovials or cryobags. The common cell banking concentration for mammalian cells is 10-15x106 cells/ mL. In this study, cell banks with different concentrations up to 200x106 cells/ mL were created and these cell banks were inoculated directly into production to test the growth profiles of the cells and the amount of monoclonal antibodies produced. The ultimate goal of this study was to reduce time, labor, cost and operational risks by using high concentration cell banks.