Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem

Today, recombinant expression of proteins is used for a variety of purposes. One of these is the production of allergens, which are vital components in allergy diagnostics. However, traditional expression systems such as ​Escherichia coli​ and ​Pichia pastoris​ might not have the capacity to express...

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Main Authors: Andersson, Pontus, Edenståhl, Selma, Eriksson, Elin, Hävermark, Tora, Nielsen, Jonas, Pihlblad, Alma
Format: Others
Language:Swedish
Published: Uppsala universitet, Institutionen för biologisk grundutbildning 2018
Subjects:
Online Access:http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-352114
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spelling ndltd-UPSALLA1-oai-DiVA.org-uu-3521142018-06-02T05:20:43ZFramtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystemsweThe future's expression systems for complex proteins : Evaluation of twelve expression systemsAndersson, PontusEdenståhl, SelmaEriksson, ElinHävermark, ToraNielsen, JonasPihlblad, AlmaUppsala universitet, Institutionen för biologisk grundutbildningUppsala universitet, Institutionen för biologisk grundutbildningUppsala universitet, Institutionen för biologisk grundutbildningUppsala universitet, Institutionen för biologisk grundutbildningUppsala universitet, Institutionen för biologisk grundutbildningUppsala universitet, Institutionen för biologisk grundutbildning2018ExpressionssystemproteinuttryckCorynebacterium glutamicumLactococcus lactisPseudomonas fluorescensPseudoalteromonas haloplanktisRalstonia eutrophaStreptomyces lividansAspergillus​Chlamydomonas reinhardtiiHansenula polymorphaLeishmania tarentolae​Physcomitrella patenssuspensionsbaserade växtcellerEngineering and TechnologyTeknik och teknologierToday, recombinant expression of proteins is used for a variety of purposes. One of these is the production of allergens, which are vital components in allergy diagnostics. However, traditional expression systems such as ​Escherichia coli​ and ​Pichia pastoris​ might not have the capacity to express all proteins of interest. Thermo Fisher, which is a leading producer of allergy tests, has requested an evaluation of different microorganisms and their capacity for heterologous protein expression in order to expand their existing toolbox of expression systems. This summary was made through a literature study, where twelve organisms were evaluated. Six eukaryotic and six prokaryotic expression systems are compared based on their ability to properly glycosylate protein, need for specific culture conditions, safety, protease activity, duration, protein yield and protein solubility. The prokaryotic systems – Corynebacterium glutamicum​ , ​Lactococcus lactis​ , ​Pseudomonas fluorescens​ , Pseudoalteromonas haloplanktis​ , ​Ralstonia eutropha​ and ​Streptomyces lividans​ – are characterized by being easy to cultivate, operating in different temperature ranges and providing relatively high yields of recombinant protein. The eukaryotic systems – ​Aspergillus fungi, the green algae ​Chlamydomonas reinhardtii​ , the yeast ​Hansenula polymorpha​ , the parasite ​Leishmania tarentolae​ , the moss ​Physcomitrella patens​ and suspension-based plant cells – all have very different morphology and properties. In comparison with the prokaryotic systems, it can be concluded that they are generally better at folding and providing the correct glycosylation patterns for mammalian and plant proteins. However, they require more time and effort to establish a competent cell line. Furthermore, the resulting protein yield is usually less than for the prokaryotic systems. The conclusion can be drawn that no expression system is perfect. The solution is a toolbox, containing various expression systems and vector systems, providing the basis for successful expression of all kinds of complex proteins. Based on the evaluation of expression systems in this review, such toolbox can be obtained. Student thesisinfo:eu-repo/semantics/bachelorThesistexthttp://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-352114application/pdfinfo:eu-repo/semantics/openAccess
collection NDLTD
language Swedish
format Others
sources NDLTD
topic Expressionssystem
proteinuttryck
Corynebacterium glutamicum
Lactococcus lactis
Pseudomonas fluorescens
Pseudoalteromonas haloplanktis
Ralstonia eutropha
Streptomyces lividans
Aspergillus
​Chlamydomonas reinhardtii
Hansenula polymorpha
Leishmania tarentolae
​Physcomitrella patens
suspensionsbaserade växtceller
Engineering and Technology
Teknik och teknologier
spellingShingle Expressionssystem
proteinuttryck
Corynebacterium glutamicum
Lactococcus lactis
Pseudomonas fluorescens
Pseudoalteromonas haloplanktis
Ralstonia eutropha
Streptomyces lividans
Aspergillus
​Chlamydomonas reinhardtii
Hansenula polymorpha
Leishmania tarentolae
​Physcomitrella patens
suspensionsbaserade växtceller
Engineering and Technology
Teknik och teknologier
Andersson, Pontus
Edenståhl, Selma
Eriksson, Elin
Hävermark, Tora
Nielsen, Jonas
Pihlblad, Alma
Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
description Today, recombinant expression of proteins is used for a variety of purposes. One of these is the production of allergens, which are vital components in allergy diagnostics. However, traditional expression systems such as ​Escherichia coli​ and ​Pichia pastoris​ might not have the capacity to express all proteins of interest. Thermo Fisher, which is a leading producer of allergy tests, has requested an evaluation of different microorganisms and their capacity for heterologous protein expression in order to expand their existing toolbox of expression systems. This summary was made through a literature study, where twelve organisms were evaluated. Six eukaryotic and six prokaryotic expression systems are compared based on their ability to properly glycosylate protein, need for specific culture conditions, safety, protease activity, duration, protein yield and protein solubility. The prokaryotic systems – Corynebacterium glutamicum​ , ​Lactococcus lactis​ , ​Pseudomonas fluorescens​ , Pseudoalteromonas haloplanktis​ , ​Ralstonia eutropha​ and ​Streptomyces lividans​ – are characterized by being easy to cultivate, operating in different temperature ranges and providing relatively high yields of recombinant protein. The eukaryotic systems – ​Aspergillus fungi, the green algae ​Chlamydomonas reinhardtii​ , the yeast ​Hansenula polymorpha​ , the parasite ​Leishmania tarentolae​ , the moss ​Physcomitrella patens​ and suspension-based plant cells – all have very different morphology and properties. In comparison with the prokaryotic systems, it can be concluded that they are generally better at folding and providing the correct glycosylation patterns for mammalian and plant proteins. However, they require more time and effort to establish a competent cell line. Furthermore, the resulting protein yield is usually less than for the prokaryotic systems. The conclusion can be drawn that no expression system is perfect. The solution is a toolbox, containing various expression systems and vector systems, providing the basis for successful expression of all kinds of complex proteins. Based on the evaluation of expression systems in this review, such toolbox can be obtained.
author Andersson, Pontus
Edenståhl, Selma
Eriksson, Elin
Hävermark, Tora
Nielsen, Jonas
Pihlblad, Alma
author_facet Andersson, Pontus
Edenståhl, Selma
Eriksson, Elin
Hävermark, Tora
Nielsen, Jonas
Pihlblad, Alma
author_sort Andersson, Pontus
title Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
title_short Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
title_full Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
title_fullStr Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
title_full_unstemmed Framtidens expressionssystem för svåruttryckta proteiner : Utvärdering av tolv expressionssystem
title_sort framtidens expressionssystem för svåruttryckta proteiner : utvärdering av tolv expressionssystem
publisher Uppsala universitet, Institutionen för biologisk grundutbildning
publishDate 2018
url http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-352114
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