Production of Erythropoietin in Pichia

PastorisEporis – Team 15

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Team Eporis

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Abby LeistraStephen TubergenZion Lee Nick Giles

StructureReview Process ModelCapacity Business Model Future Steps

Background: What is Erythropoietin?

•Erythropoietin (EPO) is a glycoprotein hormone•Naturally made in kidneys•Promotes red blood cell production•Treats anemia caused by• Chronic kidney disease

• Cancer chemotherapy

• HIV therapy

• $4 Billion US Market in 2013

glycam.org

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Problem

• Chinese Hamster Ovary (CHO) Cells• Slow-growing• Many purification steps

nikon2.magnet.fsu.edu

Solution

2013.igem.org

• Pichia Pastoris Cells• Fast-growing• Easy purification of protein

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EPO Structure

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Nett, J. H.; Gomathinayagam, S.; et. al. J. Biotechnol. 2011, 157, 198-206.

•Polyethylene Glycol (PEG) • Large

• Negatively charged

Capacity Estimate

• How much? How big?

• Depends on:

• Market Volume

• Yeast Productivity

• Separation Recovery

en.wikipedia.org

• Bottom Line: 169 g EPO/yr → 8500 L/yr

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Process Model -PFD

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http://www.esslab.com/Hamilton/sensors/sensor_applications/bio.html

Process Model -Bioreactor

• Key Considerations• Oxygen Distribution• Impeller Shear Force

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http://2010.igem.org/File:UCL-BIOREACTSRS.png.

Process Model - Filtration

•Ultrafiltration

•Microfiltration

Houp, R. C. J. Validation Tech. 2009, 40-45.

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Business Model

•Three different alternatives• Start –up• Contracting • Leasing

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stcaustralia.org

Future Steps

•Provide detail to the process model• Calculating flow rates in and out of major process units• Determining process subunits

•Evaluate alternatives• Business model• Process units

•Optimize selected process model

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Questions?