http://publicationslist.org/data/sophia.hober/atom.xml Sophia Hoberhttp://publicationslist.org/sophia.hober$basepathfavicon.icoRecent additions to Sophia Hober's PublicationsList.org pagehttp://publicationslist.org/publications.png2009-05-20T08:11:16Z http://publicationslist.org/sophia.hober/refid52 2009-05-20T08:10:19Z

There is a need for protein-specific affinity reagents to explore the gene products encoded by the genome. Recently, systematic efforts to generate validated affinity reagents on a whole human proteome level have been initiated. There are several issues for such efforts, including choice of antigen, type of affinity reagent, and the subsequent validation of the generated protein-specific binders. ...<br/><br/>Mathias Uhlén, Sophia Hober (2009) <i>J Mol Recognit</i> <i></i> <i></i> 22: 2 57-64<br/>

http://publicationslist.org/sophia.hober/refid58 2009-05-20T08:10:19Z

Various approaches for removal of high-abundance components in body fluids are currently available. While most methods are constructed for plasma depletion, there is a need for body-fluid-specific strategies. The aim of the present study was to design an affinity matrix suitable for the depletion of high-abundance proteins in CSF (cerebrospinal fluid). Hence, molecules with specific affinity towar...<br/><br/>Margareta Ramström, Aida Zuberovic, Caroline Grönwall, Jörg Hanrieder, Jonas Bergquist, Sophia Hober (2009) <i>Biotechnol Appl Biochem</i> <i></i> <i></i> 52: Pt 2 159-166<br/>

http://publicationslist.org/sophia.hober/refid53 2009-05-20T08:10:19Z

A need exists for mapping the protein profiles in the human brain both during normal and disease conditions. Here, we have studied 800 antibodies generated towards human proteins as part of a Human Protein Atlas program and investigated their suitability for detailed analysis of various levels of a rat brain using immuno-based methods. In this way, the parallel, rather limited analysis of the huma...<br/><br/> Mulder, Björling, Jonasson, Wernérus, Hober, Hökfelt, Uhlén (2009) <i>Mol Cell Proteomics</i> <i></i> <i></i> : <br/>

http://publicationslist.org/sophia.hober/refid54 2009-05-20T08:10:19Z

A completely automated procedure for the purification and desalting of proteins with a polyhistidine purification tag prior to mass spectrometry analysis is presented. The system is ideal for rapid quality control and optimization studies and it provides researchers with a straightforward, reliable tool for studies of recombinant proteins. Forty-eight samples can be prepared within 4.5h and only s...<br/><br/>Johanna Steen, Margareta Ramström, Mathias Uhlén, Sophia Hober, Jenny Ottosson (2009) <i>J Chromatogr A</i> <i></i> <i></i> 1216: 20 4457-4464<br/>

http://publicationslist.org/sophia.hober/refid55 2009-05-20T08:10:19Z

Antibody-based proteomics efforts depend on validated antibodies to ensure correct annotation of analyzed proteins. We have previously argued that a low sequence identity to other proteins is a key feature for antigens used in antibody generation. Thus, a major challenge for whole-proteome studies is how to address families of highly sequence related proteins within the context of generating speci...<br/><br/>K Larsson, C Eriksson, J M Schwenk, L Berglund, K Wester, M Uhlén, S Hober, H Wernérus (2009) <i>J Immunol Methods</i> <i></i> <i></i> 342: 1-2 20-32<br/>

http://publicationslist.org/sophia.hober/refid56 2009-05-20T08:10:19Z

The demand for high-throughput recombinant protein production has markedly increased with the increased activity in the field of proteomics. Within the Human Protein Atlas project recombinantly produced human protein fragments are used for antibody production. Here we describe how the protein expression and purification protocol has been optimized in the project to allow for handling of nearly 300...<br/><br/>Hanna Tegel, Johanna Steen, Anna Konrad, Hero Nikdin, Katarina Pettersson, Maria Stenvall, Samuel Tourle, Ulla Wrethagen, LanLan Xu, Louise Yderland, Mathias Uhlén, Sophia Hober, Jenny Ottosson (2009) <i>Biotechnol J</i> <i></i> <i></i> 4: 1 51-57<br/>

http://publicationslist.org/sophia.hober/refid1 2008-08-13T12:59:25Z

Here we report the development of a publicly available Web-based analysis tool for exploring proteins expressed in a tissue- or cancer-specific manner. The search queries are based on the human tissue profiles in normal and cancer cells in the Human Protein Atlas portal and rely on the individual annotation performed by pathologists of images representing immunohistochemically stained tissue secti...<br/><br/>Erik Björling, Cecilia Lindskog, Per Oksvold, Jerker Linné, Caroline Kampf, Sophia Hober, Mathias Uhlén, Fredrik Pontén (2008) <i>Mol Cell Proteomics</i> <i></i> <i></i> 7: 5 825-844<br/>

http://publicationslist.org/sophia.hober/refid47 2008-05-05T10:12:23Z

Currently one of the most challenging tasks in biological and medical research is to explore and understand the function of all proteins encoded by the genome of an organism. A systematic approach based on the genome sequences is feasible because the full genome of many organisms presently is available and many more are underway. For the production of expression atlases different strategies are us...<br/><br/>S Hober, M Uhlén (2008) <i>Curr Opin Biotechnol</i> <i></i> <i></i> 19: 1 30-35<br/>

http://publicationslist.org/sophia.hober/refid49 2008-05-05T10:14:52Z

Various approaches for removal of high abundant components in body fluids are currently available. While most methods are constructed for plasma depletion, there is a need for body fluid specific strategies. The aim of this project was to design an affinity matrix suitable for the depletion of high-abundant proteins in cerebrospinal fluid (CSF). Hence, molecules with specific affinity towards prot...<br/><br/> Ramström, Zuberovic, Grönwall, Hanrieder, Bergquist, Hober (2008) <i>Biotechnol Appl Biochem</i> <i></i> <i></i> : <br/>

http://publicationslist.org/sophia.hober/refid51 2009-05-20T08:10:19Z

An attractive path forward in proteomics is to experimentally annotate the human protein complement of the genome in a genecentric manner. Using antibodies, it might be possible to design protein-specific probes for a representative protein from every protein-coding gene and to subsequently use the antibodies for systematical analysis of cellular distribution and subcellular localization of protei...<br/><br/>Lisa Berglund, Erik Björling, Per Oksvold, Linn Fagerberg, Anna Asplund, Cristina Al-Khalili Szigyarto, Anja Persson, Jenny Ottosson, Henrik Wernérus, Peter Nilsson, Emma Lundberg, Asa Sivertsson, Sanjay Navani, Kenneth Wester, Caroline Kampf, Sophia Hober, Fredrik Pontén, Mathias Uhlén (2008) <i>Mol Cell Proteomics</i> <i></i> <i></i> 7: 10 2019-2027<br/>

http://publicationslist.org/sophia.hober/refid50 2009-05-20T08:10:19Z

Generation of monospecific antibodies (msAbs) (multiepitope) through affinity purification of polyclonal antisera is a plausible strategy for high-throughput production of affinity reagents toward large sets of proteins. These antibodies are generated using readily accessible gene sequence information from publicly available databases. The resulting antibodies have the potential to be used in a va...<br/><br/>Linda Paavilainen, Henrik Wernérus, Peter Nilsson, Mathias Uhlén, Sophia Hober, Kenneth Wester, Fredrik Pontén (2008) <i>Appl Immunohistochem Mol Morphol</i> <i></i> <i></i> 16: 5 493-502<br/>

http://publicationslist.org/sophia.hober/refid57 2009-05-20T08:10:19Z

We have identified a gene, denoted PttMAP20, which is strongly up-regulated during secondary cell wall synthesis and tightly coregulated with the secondary wall-associated CESA genes in hybrid aspen (Populus tremula x tremuloides). Immunolocalization studies with affinity-purified antibodies specific for PttMAP20 revealed that the protein is found in all cell types in developing xylem and that it ...<br/><br/>Alex S Rajangam, Manoj Kumar, Henrik Aspeborg, Gea Guerriero, Lars Arvestad, Podjamas Pansri, Christian J-L Brown, Sophia Hober, Kristina Blomqvist, Christina Divne, Ines Ezcurra, Ewa Mellerowicz, Björn Sundberg, Vincent Bulone, Tuula T Teeri (2008) <i>Plant Physiol</i> <i></i> <i></i> 148: 3 1283-1294<br/>

http://publicationslist.org/sophia.hober/refid6 2007-11-05T07:59:16Z

With the completion of the human genome project (HUGO) during recent years, gene function, protein abundance and expression patterns in tissues and cell types have emerged as central areas for the scientific community. A mapped human proteome will extend the value of the genome sequence and large-scale efforts aiming at elucidating protein localization, abundance and function are invaluable for bi...<br/><br/>Ronny Falk, Margareta Ramström, Stefan Ståhl, Sophia Hober (2007) <i>Biomol Eng</i> <i></i> <i></i> 24: 2 155-168<br/>

http://publicationslist.org/sophia.hober/refid7 2007-11-05T07:59:16Z

RFP2, a gene frequently lost in various malignancies, encodes a protein with RING finger, B-box, and coiled-coil domains that belongs to the RBCC/TRIM family of proteins. Here we demonstrate that Rfp2 is an unstable protein with auto-polyubiquitination activity in vivo and in vitro, implying that Rfp2 acts as a RING E3 ubiquitin ligase. Consequently, Rfp2 ubiquitin ligase activity is dependent on ...<br/><br/>Mikael Lerner, Martin Corcoran, Diana Cepeda, Michael L Nielsen, Roman Zubarev, Fredrik Pontén, Mathias Uhlén, Sophia Hober, Dan Grandér, Olle Sangfelt (2007) <i>Mol Biol Cell</i> <i></i> <i></i> 18: 5 1670-1682<br/>

http://publicationslist.org/sophia.hober/refid5 2007-11-05T07:59:16Z

Completion of the Human Genome Project and recent developments in proteomics make it possible to systematically generate affinity reagents to a large portion of the proteome. Recently an antibody-based human protein atlas covering many organs including four areas of the brain has been released (www.proteinatlas.org). Due to the heterogeneity, size, and availability of tissue a more thorough analys...<br/><br/>J Mulder, H Wernérus, T-J Shi, F Pontén, S Hober, M Uhlén, T Hökfelt (2007) <i>Neuroscience</i> <i></i> <i></i> 146: 4 1689-1703<br/>

http://publicationslist.org/sophia.hober/refid8 2007-11-05T07:59:16Z

Staphylococcal protein A (SPA) is one of the first discovered immunoglobulin binding molecules and has been extensively studied during the past decades. Due to its affinity to immunoglobulins, SPA has found widespread use as a tool in the detection and purification of antibodies and the molecule has been further developed to one of the most employed affinity purification systems. Interestingly, a ...<br/><br/>Sophia Hober, Karin Nord, Martin Linhult (2007) <i>J Chromatogr B Analyt Technol Biomed Life Sci</i> <i></i> <i></i> 848: 1 40-47<br/>

http://publicationslist.org/sophia.hober/refid4 2007-11-05T07:59:16Z

A high-throughput protein purification strategy using the polycationic Z(basic) tag has been developed. In order for the strategy to be useful both for soluble and less soluble proteins, a denaturating agent, urea, was used in all purification steps. First, four target proteins were genetically fused to the purification tag, Z(basic). These protein constructs were purified by cation exchange chrom...<br/><br/>Tove Alm, Johanna Steen, Jenny Ottosson, Sophia Hober (2007) <i>Biotechnol J</i> <i></i> <i></i> 2: 6 709-716<br/>

http://publicationslist.org/sophia.hober/refid2 2008-02-06T14:33:21Z

An Affibody (Affibody) ligand with specific binding to human transferrin was selected by phage display technology from a combinatorial protein library based on the staphylococcal protein A (SpA)-derived Z domain. Strong and selective binding of the selected Affibody ligand to transferrin was demonstrated using biosensor technology and dot blot analysis. Impressive specificity was demonstrated as t...<br/><br/>Caroline Grönwall, Anna Sjöberg, Margareta Ramström, Ingmarie Höidén-Guthenberg, Sophia Hober, Per Jonasson, Stefan Ståhl (2007) <i>Biotechnol J</i> <i></i> <i></i> 2: 11 1389-1398<br/>

http://publicationslist.org/sophia.hober/refid3 2007-11-05T07:59:16Z

A positively charged protein domain, Z(basic), can be used as a general purification tag to achieve efficient recovery of recombinantly produced target proteins using cation-exchange chromatography. To construct a protein domain usable for ion-exchange chromatography, the surface of protein Z was engineered to be highly charged, which allowed for selective capture of target proteins on a cation-ex...<br/><br/>My Hedhammar, Sophia Hober (2007) <i>J Chromatogr A</i> <i></i> <i></i> 1161: 1-2 22-28<br/>

http://publicationslist.org/sophia.hober/refid14 2007-11-05T07:59:16Z

One of the key steps in high-throughput protein production is protein purification. A newly developed high-yield protein purification and isolation method for laboratory scale use is presented. This procedure allows fully automated purification of up to 60 cell lysates with milligram yields of pure recombinant protein in 18.5h. The method is based on affinity chromatography and has been set up on ...<br/><br/>Johanna Steen, Mathias Uhlén, Sophia Hober, Jenny Ottosson (2006) <i>Protein Expr Purif</i> <i></i> <i></i> 46: 2 173-178<br/>

http://publicationslist.org/sophia.hober/refid15 2007-11-05T07:59:16Z

A strategy for efficient cleavage of fusion proteins using an immobilised protease has been developed. Protease 3C from coxsackie virus was recombinantly produced in Escherichia coli and covalently immobilised onto a solid support. Thereafter, Z(basic) tagged fusion proteins, with a specific cleavage sequence between the domains, were flown through the proteolytic column and circulated until compl...<br/><br/>M Hedhammar, H R Jung, S Hober (2006) <i>Protein Expr Purif</i> <i></i> <i></i> 47: 2 422-426<br/>

http://publicationslist.org/sophia.hober/refid17 2007-11-05T07:59:16Z

BACKGROUND: Pyrosequencing is a DNA sequencing technology based on the sequencing-by-synthesis principle. METHODS: The technique is built on a 4-enzyme real-time monitoring of DNA synthesis by bioluminescence using a cascade that upon nucleotide incorporation ends in a detectable light signal (bioluminescence). The detection system is based on the pyrophosphate released when a nucleotide is introd...<br/><br/>Afshin Ahmadian, Maria Ehn, Sophia Hober (2006) <i>Clin Chim Acta</i> <i></i> <i></i> 363: 1-2 83-94<br/>

http://publicationslist.org/sophia.hober/refid13 2007-11-05T07:59:16Z

Mantle cell lymphoma (MCL) is an aggressive lymphoid malignancy for which better treatment strategies are needed. To identify potential diagnostic and therapeutic targets, a signature consisting of MCL-associated genes was selected based on a comprehensive gene expression analysis of malignant and normal B cells. The corresponding protein epitope signature tags were identified and used to raise mo...<br/><br/>Sara Ek, Ulrika Andréasson, Sophia Hober, Caroline Kampf, Fredrik Pontén, Mathias Uhlén, Hartmut Merz, Carl A K Borrebaeck (2006) <i>Mol Cell Proteomics</i> <i></i> <i></i> 5: 6 1072-1081<br/>

http://publicationslist.org/sophia.hober/refid11 2007-11-05T07:59:16Z

A new and flexible technology for high throughput analysis of antibody specificity and affinity is presented. The method is based on microfluidics and takes advantage of compact disks (CDs) in which the centrifugal force moves fluids through microstructures containing immobilized metal affinity chromatography columns. Analyses are performed as a sandwich assay, where antigen is captured to the col...<br/><br/>Cecilia Eriksson, Charlotta Agaton, Rikard Kånge, Mårten Sundberg, Peter Nilsson, Bo Ek, Mathias Uhlén, Magnus Gustafsson, Sophia Hober (2006) <i>J Proteome Res</i> <i></i> <i></i> 5: 7 1568-1574<br/>

http://publicationslist.org/sophia.hober/refid9 2007-11-05T07:59:16Z

Monospecific antibodies dfdfdfdf (msAbs) generated through antigen specific purification of polyclonal antisera are valuable tools in proteome analyses. However, proteome wide generation of msAbs would require extensive immunization programs. Therefore, it would be desirable to develop efficient immunization and purification methods to reduce the number of animals needed for such antibody-based re...<br/><br/>Karin Larsson, Kenneth Wester, Peter Nilsson, Mathias Uhlén, Sophia Hober, Henrik Wernérus (2006) <i>J Immunol Methods</i> <i></i> <i></i> 315: 1-2 110-120<br/>

http://publicationslist.org/sophia.hober/refid12 2007-11-05T07:59:16Z

The Human Protein Atlas is a comprehensive database that provides the protein expression profiles for a large number of human proteins, presented as immunohistological images from most human tissues. This review provides an overview of the contents of the atlas, discusses the project strategy and highlights the importance of open access for data validation and quality. Essential procedures that ar...<br/><br/>Anja Persson, Sophia Hober, Mathias Uhlén (2006) <i>Curr Opin Mol Ther</i> <i></i> <i></i> 8: 3 185-190<br/>

http://publicationslist.org/sophia.hober/refid10 2007-11-05T07:59:16Z

A strategy for purification of inclusion body-forming proteins is described, in which the positively charged domain Z(basic) is used as a fusion partner for capture of denatured proteins on a cation exchange column. It is shown that the purification tag is selective under denaturing conditions. Furthermore, the new strategy for purification of proteins from inclusion bodies is compared with the co...<br/><br/>My Hedhammar, Tove Alm, Torbjörn Gräslund, Sophia Hober (2006) <i>Biotechnol J</i> <i></i> <i></i> 1: 2 187-196<br/>

http://publicationslist.org/sophia.hober/refid20 2007-11-05T07:59:16Z

A high throughput method for screening of protein expression is described. By using a flow cytometer, levels of both soluble and precipitated protein can simultaneously be assessed in vivo. Protein fragments were fused to the N-terminus of enhanced GFP and the cell samples were analysed using a flow cytometer. Data concerning whole cell fluorescence and light scattering was collected. The whole ce...<br/><br/>My Hedhammar, Maria Stenvall, Rosa Lönneborg, Olof Nord, Olle Sjölin, Hjalmar Brismar, Mathias Uhlén, Jenny Ottosson, Sophia Hober (2005) <i>J Biotechnol</i> <i></i> <i></i> 119: 2 133-146<br/>

http://publicationslist.org/sophia.hober/refid21 2007-11-05T07:59:16Z

Significant efforts are put into the design of large-scale purification processes of proteins due to great demands regarding cost efficiency and safety. In order to design an effective purification scheme the unit operations need to be reduced to a minimum. In this review we are discussing proteinaceous ligands as well as small synthetic mimics for use in affinity chromatography for large-scale ap...<br/><br/>Martin Linhult, Susanne Gülich, Sophia Hober (2005) <i>Protein Pept Lett</i> <i></i> <i></i> 12: 4 305-310<br/>

http://publicationslist.org/sophia.hober/refid22 2007-11-05T07:59:16Z

The effect of changes in substrate feed rate during fedbatch cultivation was investigated with respect to soluble protein formation and transport of product to the periplasm in Escherichia coli. Production was transcribed from the P(malK) promoter; and the cytoplasmic part of the production was compared with production from the P(lacUV5) promoter. The fusion protein product, Zb-MalE, was at all ti...<br/><br/>Maria Boström, Katrin Markland, Anna Maria Sandén, My Hedhammar, Sophia Hober, Gen Larsson (2005) <i>Appl Microbiol Biotechnol</i> <i></i> <i></i> 68: 1 82-90<br/>

http://publicationslist.org/sophia.hober/refid16 2007-11-05T07:59:16Z

A great need exists for the systematic generation of specific antibodies to explore the human proteome. Here, we show that antibodies specific to human proteins can be generated in a high-throughput manner involving stringent affinity purification using recombinant protein epitope signature tags (PrESTs) as immunogens and affinity-ligands. The specificity of the generated affinity reagents, here c...<br/><br/>Peter Nilsson, Linda Paavilainen, Karin Larsson, Jenny Odling, Mårten Sundberg, Ann-Catrin Andersson, Caroline Kampf, Anja Persson, Cristina Al-Khalili Szigyarto, Jenny Ottosson, Erik Björling, Sophia Hober, Henrik Wernérus, Kenneth Wester, Fredrik Pontén, Mathias Uhlen (2005) <i>Proteomics</i> <i></i> <i></i> 5: 17 4327-4337<br/>

http://publicationslist.org/sophia.hober/refid18 2007-11-05T07:59:16Z

Antibody-based proteomics provides a powerful approach for the functional study of the human proteome involving the systematic generation of protein-specific affinity reagents. We used this strategy to construct a comprehensive, antibody-based protein atlas for expression and localization profiles in 48 normal human tissues and 20 different cancers. Here we report a new publicly available database...<br/><br/>Mathias Uhlén, Erik Björling, Charlotta Agaton, Cristina Al-Khalili Szigyarto, Bahram Amini, Elisabet Andersen, Ann-Catrin Andersson, Pia Angelidou, Anna Asplund, Caroline Asplund, Lisa Berglund, Kristina Bergström, Harry Brumer, Dijana Cerjan, Marica Ekström, Adila Elobeid, Cecilia Eriksson, Linn Fagerberg, Ronny Falk, Jenny Fall, Mattias Forsberg, Marcus Gry Björklund, Kristoffer Gumbel, Asif Halimi, Inga Hallin, Carl Hamsten, Marianne Hansson, My Hedhammar, Görel Hercules, Caroline Kampf, Karin Larsson, Mats Lindskog, Wald Lodewyckx, Jan Lund, Joakim Lundeberg, Kristina Magnusson, Erik Malm, Peter Nilsson, Jenny Odling, Per Oksvold, Ingmarie Olsson, Emma Oster, Jenny Ottosson, Linda Paavilainen, Anja Persson, Rebecca Rimini, Johan Rockberg, Marcus Runeson, Asa Sivertsson, Anna Sköllermo, Johanna Steen, Maria Stenvall, Fredrik Sterky, Sara Strömberg, Mårten Sundberg, Hanna Tegel, Samuel Tourle, Eva Wahlund, Annelie Waldén, Jinghong Wan, Henrik Wernérus, Joakim Westberg, Kenneth Wester, Ulla Wrethagen, Lan Lan Xu, Sophia Hober, Fredrik Pontén (2005) <i>Mol Cell Proteomics</i> <i></i> <i></i> 4: 12 1920-1932<br/>

http://publicationslist.org/sophia.hober/refid48 2008-05-05T10:18:23Z

When producing and purifying recombinant proteins it is of importance to minimize the number of unit operations during the purification procedure. This is accomplished by increasing the selectivity in each step. Due to the high selectivity of affinity chromatography it has a widespread use in protein purification. However, most target proteins lack a suitable affinity ligand usable for captu...<br/><br/>Hedhammar, M., Gräslund, T., & Hober, S (2005) <i>Chemical Engineering & Technology</i> 28: 1315-1325<br/>

http://publicationslist.org/sophia.hober/refid19 2007-11-05T07:59:16Z

A high-throughput assay is described for analysis of the solubility of purified recombinant proteins. The assay is based on affinity purification of proteins in the presence of chaotropic agents followed by a dilution and incubation step to investigate the solubility in the absence of high concentrations of such agents. The assay can be performed in a 96-well format, which makes it well suited for...<br/><br/>Maria Stenvall, Johanna Steen, Mathias Uhlén, Sophia Hober, Jenny Ottosson (2005) <i>Biochim Biophys Acta</i> <i></i> <i></i> 1752: 1 6-10<br/>

http://publicationslist.org/sophia.hober/refid23 2007-11-05T07:59:16Z

A novel strategy for the highly selective purification of recombinant fusion proteins using negatively charged protein domains, which were constructed by protein design, is described. A triple alpha-helical domain of 58 amino acids was used as scaffold. Far-ultraviolet circular dichroism measurements showed that the designed domains had very low alpha-helicity in a low-conductivity environment in ...<br/><br/>M Hedhammar, T Gräslund, M Uhlén, S Hober (2004) <i>Protein Eng Des Sel</i> <i></i> <i></i> 17: 11 779-786<br/>

http://publicationslist.org/sophia.hober/refid24 2007-11-05T07:59:16Z

A high stringency protocol, suitable for systematic purification of polyclonal antibodies, is described. The procedure is designed to allow the generation of target protein-specific antibodies suitable for functional annotation of proteins. Antibodies were generated by immunization with recombinantly produced affinity-tagged target proteins. To obtain stringent recovery of the antibodies, a two-st...<br/><br/>Charlotta Agaton, Ronny Falk, Ingmarie Höidén Guthenberg, Lovisa Göstring, Mathias Uhlén, Sophia Hober (2004) <i>J Chromatogr A</i> <i></i> <i></i> 1043: 1 33-40<br/>

http://publicationslist.org/sophia.hober/refid26 2007-11-05T07:59:16Z

Mutation detection and single-nucleotide polymorphism genotyping require screening of large samples of materials and therefore the importance of high-throughput DNA analysis techniques is significant. Pyrosequencing is a four-enzyme bioluminometric DNA sequencing technology based on the sequencing-by-synthesis principle. Currently, the technique is limited to simultaneous analysis of 96 or 384 sam...<br/><br/>Maria Ehn, Nader Nourizad, Kristina Bergström, Afshin Ahmadian, Pål Nyrén, Joakim Lundeberg, Sophia Hober (2004) <i>Anal Biochem</i> <i></i> <i></i> 329: 1 11-20<br/>

http://publicationslist.org/sophia.hober/refid25 2007-11-05T07:59:16Z

Protein-protein interactions play crucial roles in various biological pathways and functions. Therefore, the characterization of protein levels and also the network of interactions within an organism would contribute considerably to the understanding of life. The availability of the human genome sequence has created a range of new possibilities for biomedical research. A crucial challenge is to ut...<br/><br/>Charlotta Agaton, Mathias Uhlén, Sophia Hober (2004) <i>Electrophoresis</i> <i></i> <i></i> 25: 9 1280-1288<br/>

http://publicationslist.org/sophia.hober/refid27 2007-11-05T07:59:16Z

Staphylococcal protein A (SPA) is a cell surface protein expressed by Staphylococcus aureus. It consists of five repetitive domains. The five SPA-domains show individual interaction to the Fc-fragment as well as certain Fab-fragments of immunoglobulin G (IgG) from most mammalian species. Due to the high affinity and selectivity of SPA, it has a widespread use as an affinity ligand for capture and ...<br/><br/>Martin Linhult, Susanne Gülich, Torbjörn Gräslund, Annelie Simon, Martin Karlsson, Anna Sjöberg, Karin Nord, Sophia Hober (2004) <i>Proteins</i> <i></i> <i></i> 55: 2 407-416<br/>

http://publicationslist.org/sophia.hober/refid30 2007-11-05T07:59:16Z

ATP-diphosphohydrolase (apyrase) catalyzes the hydrolysis of phosphoanhydride bonds of nucleoside tri- and di-phosphates in the presence of divalent cations. This enzyme has broad substrate specificity for nucleotides, which makes it an ideal enzyme for different biotechnical applications, such as DNA sequencing and platelet-aggregation inhibition. The only commercially available apyrase is isolat...<br/><br/>Nader Nourizad, Maria Ehn, Baback Gharizadeh, Sophia Hober, Pål Nyrén (2003) <i>Protein Expr Purif</i> <i></i> <i></i> 27: 2 229-237<br/>

http://publicationslist.org/sophia.hober/refid28 2007-11-05T07:59:16Z

Alkaline conditions are generally preferred for sanitization of chromatography media by cleaning-in-place (CIP) protocols in industrial biopharmaceutical processes. The use of such rigorous conditions places stringent demands on the stability of ligands intended for use in affinity chromatography. Here, we describe efforts to meet these requirements for a divalent proteinaceous human serum albumin...<br/><br/>Martin Linhult, Susanne Gülich, Torbjörn Gräslund, Per-Ake Nygren, Sophia Hober (2003) <i>Protein Eng</i> <i></i> <i></i> 16: 12 1147-1152<br/>

http://publicationslist.org/sophia.hober/refid29 2007-11-05T07:59:16Z

A novel, improved dual bacterial-expression system, designed for large-scale generation of high-quality polyclonal antibody preparations intended for proteomics research, is presented. The concept involves parallel expression of cDNA-encoded proteins, as a fusion with two different tags in two separate vector systems. Both systems enable convenient blotting procedures for expression screening on c...<br/><br/>Ronny Falk, Charlotta Agaton, Eva Kiesler, Shaobo Jin, Lars Wieslander, Neus Visa, Sophia Hober, Stefan Ståhl (2003) <i>Biotechnol Appl Biochem</i> <i></i> <i></i> 38: Pt 3 231-239<br/>

http://publicationslist.org/sophia.hober/refid35 2007-11-05T07:59:16Z

Streptococcal protein G (SpG) is a bacterial cell surface receptor exhibiting affinity to both human immunoglobulin (IgG) and human serum albumin (HSA). Interestingly, the serum albumin and immunoglobulin-binding activities have been shown to reside at functionally and structurally separated receptor domains. The binding domain of the HSA-binding part has been shown to be a 46-residue triple alpha...<br/><br/>Martin Linhult, Hans Kaspar Binz, Mathias Uhlén, Sophia Hober (2002) <i>Protein Sci</i> <i></i> <i></i> 11: 2 206-213<br/>

http://publicationslist.org/sophia.hober/refid36 2007-11-05T07:59:16Z

We have determined the solution structure of an albumin binding domain of protein G, a surface protein of group C and G streptococci. We find that it folds into a left handed three-helix bundle similar to the albumin binding domain of protein PAB from Peptostreptococcus magnus. The two domains share 59% sequence identity, are thermally very stable, and bind to the same site on human serum albumin....<br/><br/>Maria U Johansson, Inga-Maria Frick, Hanna Nilsson, Per J Kraulis, Sophia Hober, Per Jonasson, Martin Linhult, Per-Ake Nygren, Mathias Uhlén, Lars Björck, Torbjörn Drakenberg, Sture Forsén, Mats Wikström (2002) <i>J Biol Chem</i> <i></i> <i></i> 277: 10 8114-8120<br/>

http://publicationslist.org/sophia.hober/refid34 2007-11-05T07:59:16Z

To achieve efficient recovery of recombinantly produced target proteins using cation-exchange chromatography, a novel basic protein domain is used as a purification handle. The proteolytic instability usually encountered for basic peptide tags is avoided by the use of a highly constrained alpha-helical domain based on staphylococcal protein A into which positively charged amino acids have been int...<br/><br/>Torbjörn Gräslund, Maria Ehn, Gunnel Lundin, My Hedhammar, Mathias Uhlén, Per-Ake Nygren, Sophia Hober (2002) <i>J Chromatogr A</i> <i></i> <i></i> 942: 1-2 157-166<br/>

http://publicationslist.org/sophia.hober/refid32 2007-11-05T07:59:16Z

Pyrosequencing is a four-enzyme bioluminometric DNA sequencing technique based on a DNA sequencing by synthesis principle. Currently, the technique is limited to analysis of short DNA sequences exemplified by single-nucleotide polymorphism analysis. In order to expand the field for pyrosequencing, the read length needs to be improved and efforts have been made to purify reaction components as well...<br/><br/>Maria Ehn, Afshin Ahmadian, Peter Nilsson, Joakim Lundeberg, Sophia Hober (2002) <i>Electrophoresis</i> <i></i> <i></i> 23: 19 3289-3299<br/>

http://publicationslist.org/sophia.hober/refid31 2007-11-05T07:59:16Z

Most protein-based affinity chromatography media are very sensitive towards alkaline treatment, which is a preferred method for regeneration and removal of contaminants from the purification devices in industrial applications. In a previous study, we concluded that a simple and straightforward strategy consisting of replacing asparagine residues could improve the stability towards alkaline conditi...<br/><br/>Susanne Gülich, Martin Linhult, Stefan Ståhl, Sophia Hober (2002) <i>Protein Eng</i> <i></i> <i></i> 15: 10 835-842<br/>

http://publicationslist.org/sophia.hober/refid33 2007-11-05T07:59:16Z

The highly charged domain Z(basic) can be used as a fusion partner to enhance adsorption of target proteins to cation exchanging resins at high pH-values. In this paper, we describe a strategy for purification of target proteins fused to Z(basic) at a constant physiological pH using cation exchange chromatography in an expanded bed mode. We show that two proteins, Klenow DNA polymerase and the vir...<br/><br/>Torbjörn Gräslund, My Hedhammar, Mathias Uhlén, Per Ake Nygren, Sophia Hober (2002) <i>J Biotechnol</i> <i></i> <i></i> 96: 1 93-102<br/>

http://publicationslist.org/sophia.hober/refid37 2007-11-05T07:59:16Z

Escherichia coli (E. coli) single-stranded binding protein (SSB) is a valuable protein for various biotechnical applications, such as PCR and DNA sequencing. Here we describe an efficient expression and purification scheme where the tendency of SSB to aggregate at low salt concentration and high protein concentration is avoided. The method contains fewer steps of purification and results in high p...<br/><br/>M Ehn, P Nilsson, M Uhlén, S Hober (2001) <i>Protein Expr Purif</i> <i></i> <i></i> 22: 1 120-127<br/>

http://publicationslist.org/sophia.hober/refid40 2007-11-05T07:59:16Z

One of the problems in the recovery of antibodies by affinity chromatography is the low pH, which is normally essential to elute the bound material from the column. Here, we have addressed this problem by constructing destabilized mutants of a domain analogue (domain Z) from an IgG-binding bacterial receptor, protein A. In order to destabilize the IgG-binding domain, two protein engineered variant...<br/><br/>S Gülich, M Uhlén, S Hober (2000) <i>J Biotechnol</i> <i></i> <i></i> 76: 2-3 233-244<br/>

http://publicationslist.org/sophia.hober/refid39 2007-11-05T07:59:16Z

One of the problems with a proteinaceous affinity ligand is their sensitivity to alkaline conditions. Here, we show that a simple and straightforward strategy consisting in replacing all asparagine residues with other amino acids can dramatically improve the chemical stability of a protein towards alkaline conditions. As a model, a Streptococcal albumin-binding domain (ABD) was used. The engineere...<br/><br/>S Gülich, M Linhult, P Nygren, M Uhlén, S Hober (2000) <i>J Biotechnol</i> <i></i> <i></i> 80: 2 169-178<br/>

http://publicationslist.org/sophia.hober/refid38 2007-11-05T07:59:16Z

We have created protein domains with extreme surface charge. These mutated domains allow for ion-exchange chromatography under conditions favourable for selective and efficient capture, using Escherichia coli as a host organism. The staphylococcal protein A-derived domain Z (Zwt) was used as a scaffold when constructing two mutants, Zbasic1 and Zbasic2, with high positive surface charge. Far-ultra...<br/><br/>T Gräslund, G Lundin, M Uhlén, P A Nygren, S Hober (2000) <i>Protein Eng</i> <i></i> <i></i> 13: 10 703-709<br/>

http://publicationslist.org/sophia.hober/refid42 2007-11-05T07:59:16Z

Insulin-like growth factor (IGF) I does not quantitatively form its three native disulfide bonds in the presence of 10 mM reduced and 1 mM oxidized glutathione in vitro [Hober, S. et al. (1992) Biochemistry 31, 1749-1756]. In this paper, we show (i) that both IGF-I and IGF-II are unable to form and maintain their native disulfide bonds at redox conditions that are similar to the situation in the s...<br/><br/>S Hober, J Lundström Ljung, M Uhlén, B Nilsson (1999) <i>FEBS Lett</i> <i></i> <i></i> 443: 3 271-276<br/>

http://publicationslist.org/sophia.hober/refid41 2007-11-05T07:59:16Z

The kinetic rate parameters for the interaction between a single domain analogue of staphylococcal protein A (Z) and a mouse-IgG3 monoclonal antibody (MAb) were measured in Hepes buffer with different chemical additives. Five buffer ingredients (pH, NaCl, DMSO, EDTA, and KSCN) were varied simultaneously in 16 experiments following a statistical experimental plan. The 16 buffers thus spanned a volu...<br/><br/>K Andersson, S Gülich, M Hämäläinen, P A Nygren, S Hober, M Malmqvist (1999) <i>Proteins</i> <i></i> <i></i> 37: 3 494-498<br/>

http://publicationslist.org/sophia.hober/refid43 2007-11-05T07:59:16Z

We have previously concluded that insulin-like growth factor-I (IGF-I) is thermodynamically unable to quantitatively form its disulfide bonds under reversible redox conditions in vitro. From detailed analyses it was hypothesized that the 47-52 disulfide is energetically unfavorable in the native IGF-I structure [Hober et al. (1992) Biochemistry 31, 1749-1756]. In this paper, this hypothesis has be...<br/><br/>S Hober, M Uhlén, B Nilsson (1997) <i>Biochemistry</i> <i></i> <i></i> 36: 15 4616-4622<br/>

http://publicationslist.org/sophia.hober/refid44 2007-11-05T07:59:16Z

Insulin-like growth factor I (IGF-I) is thermodynamically unable to quantitatively form its native disulfides under reversible redox conditions in vitro [Hober et al. (1992) Biochemistry 31, 1749-1756]. These results prompted the question of how IGF-I may overcome this energetic problem in its folding in vivo. Here, we report that an IGF-I precursor, IGF-I-Ea, shows disulfide-exchange folding prop...<br/><br/>S Hober, A Hansson, M Uhlén, B Nilsson (1994) <i>Biochemistry</i> <i></i> <i></i> 33: 22 6758-6761<br/>

http://publicationslist.org/sophia.hober/refid45 2007-11-05T07:59:16Z

The disulfide exchange folding properties of insulin-like growth factor I (IGF-I) have been analyzed in a redox buffer containing reduced (10 mM) and oxidized (1 mM) glutathione. Under these conditions, the 3 disulfide bridges of the 70 amino acid peptide were not quantitatively formed. Instead, five major forms of IGF-I were detected, and these components were concluded to be in equilibrium as th...<br/><br/>S Hober, G Forsberg, G Palm, M Hartmanis, B Nilsson (1992) <i>Biochemistry</i> <i></i> <i></i> 31: 6 1749-1756<br/>

http://publicationslist.org/sophia.hober/refid46 2007-11-05T07:59:16Z

Using protein engineering, a tripartite fusion protein was constructed consisting of five IgG-binding regions of protein A from Staphylococcus aureus, two IgG-binding regions of protein G from Streptococcus strain G148 and beta-galactosidase from Escherichia coli. The resulting protein lacks the serum albumin binding regions of native protein G. The fusion protein, which is a tetramer of approxima...<br/><br/>L Strandberg, S Hober, M Uhlén, S O Enfors (1990) <i>J Biotechnol</i> <i></i> <i></i> 13: 1 83-96<br/>