| 1. |
- Brofelth, Mattias, et al.
(author)
-
Site-specific photocoupling of pBpa mutated scFv antibodies for use in affinity proteomics
- 2017
-
In: Biochimica et Biophysica Acta - Proteins and Proteomics. - : Elsevier BV. - 1570-9639. ; 1865:8, s. 985-996
-
Journal article (peer-reviewed)abstract
- Recombinant antibody libraries can provide a source of renewable and high-performing binders tailored for use in affinity proteomics. In this context, the process of generating site-specific 1:1 tagging/functionalization and/or orientated surface immobilization of antibodies has, however, proved to be challenging. Hence, novel ways of generating such engineered antibodies for use in affinity proteomics could have a major impact on array performance. In this study, we have further tailored the design of human recombinant scFv antibodies for site-specific photocoupling through the use of an unnatural amino acid (UAA) and the Dock'n'Flash technology. In more detail, we have generated the 2nd generation of scFvs carrying the photoreactive UAA p-benzoyl-l-phenylalanine (pBpa). Based on key properties, such as expression levels, activity, and affinity, a preferred choice of site for pBpa, located in the beginning of the C-terminal affinity-tag, was for the first time pin-pointed. Further, the results showed that pBpa mutated antibody could be site-specifically photocoupled to free and surface immobilized β-cyclodextrin (an affinity ligand to pBpa). This paves the way for use of scFv antibodies, engineered for site-specific photochemical-based tagging, functionalization, and orientated surface immobilization, in affinity proteomics.
|
|
| 2. |
- Carney Almroth, Bethanie, 1974, et al.
(author)
-
Quantifying shedding of synthetic fibers from textiles; a source of microplastics released into the environment
- 2018
-
In: Environmental Science and Pollution Research. - : Springer Science and Business Media LLC. - 0944-1344 .- 1614-7499. ; 25:2, s. 1191-1199
-
Journal article (peer-reviewed)abstract
- Microplastics in the environment are a subject of intense research as they pose a potential threat to marine organisms. Plastic fibers from textiles have been indicated as a major source of this type of contaminant, entering the oceans via wastewater and diverse non-point sources. Their presence is also documented in terrestrial samples. In this study, the amount of microfibers shedding from synthetic textiles was measured for three materials (acrylic, nylon, polyester), knit using different gauges and techniques. All textiles were found to shed, but polyester fleece fabrics shed the greatest amounts, averaging 7360 fibers/m(-2)/L-1 in one wash, compared with polyester fabrics which shed 87 fibers/m(-2)/L-1. We found that loose textile constructions shed more, as did worn fabrics, and high twist yarns are to be preferred for shed reduction. Since fiber from clothing is a potentially important source of microplastics, we suggest that smarter textile construction, prewashing and vacuum exhaustion at production sites, and use of more efficient filters in household washing machines could help mitigate this problem.
|
|
| 3. |
- Hazarika, Pompi, et al.
(author)
-
Photopatterning of self assembled monolayers on oxide surfaces for the selective attachment of biomolecules
- 2014
-
In: Biosensors & Bioelectronics. - : Elsevier BV. - 1873-4235 .- 0956-5663. ; 53, s. 82-89
-
Journal article (peer-reviewed)abstract
- The immobilization of functional biomolecules to surfaces is a critical process for the development of biosensors for disease diagnostics. In this work we report the patterned attachment of single chain fragment variable (scFv) antibodies to the surface of metal oxides by the photodeprotection of self-assembled monolayers, using near-UV light. The photodeprotection step alters the functionality at the surface; revealing amino groups that are utilized to bind biomolecules in the exposed regions of the substrate only. The patterned antibodies are used for the detection of specific disease biomarker proteins in buffer and in complex samples such as human serum. (C) 2013 Elsevier B.V. All rights reserved.
|
|
| 4. |
- Johnsson, Richard, et al.
(author)
-
Reductive openings of benzylidene acetals. Kinetic studies of borane and alane activation by Lewis acids.
- 2008
-
In: Carbohydrate Research. - : Elsevier BV. - 1873-426X .- 0008-6215. ; 343, s. 2997-3000
-
Journal article (peer-reviewed)abstract
- The reaction kinetics for a number of reductive openings of methyl 2,3-di-O-benzyl-4,6-O-benzylidene-alpha-d-glucopyranoside have been investigated. Openings to give free HO-6 (using BH(3).THF-AlCl(3)-THF or LiAlH(4)-AlCl(3)-Et(2)O) follow first order kinetics, while reactions yielding free HO-4 (using BH(3).NMe(3)-AlCl(3)-THF or BH(3).NMe(3)-BF(3).OEt(2)-THF) follow higher order kinetics. The addition of water to the BH(3).NMe(3)-AlCl(3)-THF results in faster reactions. The BH(3).SMe(2)-AlCl(3)-THF system constitutes a borderline case, yielding both free HO-6 (by a first order reaction) and free HO-4 (by a higher order reaction). These results correlate well with the concept of regioselectivity by activation of borane complexes.
|
|
| 5. |
- Lygum, Victoria Linn, et al.
(author)
-
Greenspace as Workplace : Benefits, Challenges and Essentialities in the Physical Environment.
- 2023
-
In: International Journal of Environmental Research and Public Health. - : MDPI. - 1661-7827 .- 1660-4601. ; 20:17
-
Journal article (peer-reviewed)abstract
- There is a scarcity of knowledge regarding the potential benefits of human-nature contact within the context of working life. Even more limited is the research that focuses on working outdoors and the setting in which it takes place. This study aimed to obtain insight into key aspects of the physical environment relevant for the experienced benefits and challenges of workers exploring office work outdoors. We conducted interviews with key informants as well as photo registration and mapping of the different green spaces in the environments of six small or medium-sized workplaces. The information gathered was used as background knowledge for exploratory qualitative interviews, which were conducted while walking in natural settings chosen by the interviewees. With a landscape architectural perspective and an inductive approach, we explored employees' experiences of bringing office work outdoors. The following themes emerged: 'Simplicity,' 'Safeness', 'Comfort', and 'Contact with Nature' were experienced as key aspects in relation to the physical environment, whereas 'Sociality', 'Well-being', and 'Functioning' stood out as the main benefits and, 'Digital dependency' and 'Illegitimacy' as challenges to overcome. Based on the identification of potential benefits and their prerequisites, we propose implications for practice and research that can be useful when focusing on bringing office work outdoors.
|
|
| 6. |
- Petersson, Linn, et al.
(author)
-
Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer.
- 2014
-
In: Nanotechnology. - : IOP Publishing. - 0957-4484 .- 1361-6528. ; 25:27
-
Journal article (peer-reviewed)abstract
- Miniaturized (Ø 10 μm), multiplexed (>5-plex), and high-density (>100 000 spots cm(-2)) antibody arrays will play a key role in generating protein expression profiles in health and disease. However, producing such antibody arrays is challenging, and it is the type and range of available spotters which set the stage. This pilot study explored the use of a novel microspotting tool, Bioplume(TM)-consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels-to produce miniaturized, multiplexed, and high-density planar recombinant antibody arrays for protein expression profiling which targets crude, directly labelled serum. The results demonstrated that 16-plex recombinant antibody arrays could be produced-based on miniaturized spot features (78.5 um(2), Ø 10 μm) at a 7-125-times increased spot density (250 000 spots cm(-2)), interfaced with a fluorescent-based read-out. This prototype platform was found to display adequate reproducibility (spot-to-spot) and an assay sensitivity in the pM range. The feasibility of the array platform for serum protein profiling was outlined.
|
|
| 7. |
- Petersson, Linn, et al.
(author)
-
Miniaturization of multiplexed planar recombinant antibody arrays for serum protein profiling.
- 2014
-
In: Bioanalysis. - : Future Science Ltd. - 1757-6180 .- 1757-6199. ; 6:9, s. 1175-1185
-
Journal article (peer-reviewed)abstract
- Background: Antibody-based microarrays are a developing tool for high-throughput proteomics in health and disease. However, in order to enable global proteome profiling, novel miniaturized high-density antibody array formats must be developed. Results: In this proof-of-concept study, we have designed a miniaturized planar recombinant (single-chain Fragment variable). antibody array technology platform for multiplexed profiling of non-fractionated, directly labelled serum samples. The size of the individual spot features was reduced 225-times (78.5 μm(2)/spot) and the array density was increased 19-times (38,000 spots/cm(2)). These miniaturized, multiplexed arrays were produced, using a desktop nanofabrication system based on dip-pen nanolithography technology, and interfaced with a high-resolution fluorescent-based scanner. The reproducibility, sensitivity, specificity, and applicability of the set-up were demonstrated by profiling a set of well-characterized serum samples. Conclusion: The designed antibody array platform opens up new possibilities for large-scale, multiplex profiling of crude proteomes in a miniaturized fashion.
|
|
| 8. |
- Petersson, Linn, et al.
(author)
-
Molecular design of recombinant scFv antibodies for site-specific photocoupling to β-cyclodextrin in solution and onto solid support.
- 2014
-
In: Biochimica et Biophysica Acta - Proteins and Proteomics. - : Elsevier BV. - 1570-9639. ; 1844:12, s. 2164-2173
-
Journal article (peer-reviewed)abstract
- The ability to design and tailor-make antibodies to meet the biophysical demands required by the vast range of current and future antibody-based applications within biotechnology and biomedicine will be essential. In this proof-of-concept study, we have for the first time tailored human recombinant scFv antibodies for site-specific photocoupling through the use of an unnatural amino acid (UAA) and the dock'n'flash technology. In more detail, we have successfully explored the possibility to expand the genetic code of E. coli and introduced the photoreactive UAA p-benzoyl-L-phenylalanine (pBpa), and showed that the mutated scFv antibody could be expressed in E. coli with retained structural and functional properties, as well as binding affinity. The pBpa group was then used for affinity capture of the mutated antibody by β-cyclodextrin (β-CD), which provided the hydrogen atoms to be abstracted in the subsequent photocoupling process upon irradiation at 365nm. The results showed that the pBpa mutated antibody could be site-specifically photocoupled to free and surface (array) immobilized β-CD. Taken together, this paves the way for novel means of tailoring recombinant scFv antibodies for site-specific photochemical-based tagging, functionalization and immobilization in numerous applications.
|
|
| 9. |
- Petersson, Linn, et al.
(author)
-
Multiplexing of miniaturized planar antibody arrays for serum protein profiling - a biomarker discovery in SLE nephritis.
- 2014
-
In: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189. ; 14:11, s. 1931-1942
-
Journal article (peer-reviewed)abstract
- In the quest to decipher disease-associated biomarkers, miniaturized and multiplexed antibody arrays may play a central role in generating protein expression profiles, or protein maps, of crude serum samples. In this conceptual study, we explored a novel, 4-times larger pen design, enabling us to, in a unique manner, simultaneously print 48 different reagents (antibodies) as individual 78.5 μm(2) (10 μm in diameter) sized spots at a density of 38 000 spots cm(-2) using dip-pen nanolithography technology. The antibody array set-up was interfaced with a high-resolution fluorescent-based scanner for sensitive sensing. The performance and applicability of this novel 48-plex recombinant antibody array platform design was demonstrated in a first clinical application targeting SLE nephritis, a severe chronic autoimmune connective tissue disorder, as the model disease. To this end, crude, directly biotinylated serum samples were targeted. The results showed that the miniaturized and multiplexed array platform displayed adequate performance, and that SLE-associated serum biomarker panels reflecting the disease process could be deciphered, outlining the use of miniaturized antibody arrays for disease proteomics and biomarker discovery.
|
|
| 10. |
- Petersson, Linn
(author)
-
Tailoring of Antibody Arrays - Technical Advances Towards Global Proteome Analysis
- 2014
-
Doctoral thesis (other academic/artistic)abstract
- Abstract The number of cancer and autoimmune disease diagnoses increases in our society, which place high demands on the health care system. Being able to tailor treatments for each individual patient is called "Personalized medicine", and could revolutionize the care in the future. This type of medicine requires biomarkers that could be used for diagnosis, prognosis and choice of therapy. In our group, we have developed an antibody microarray discovery tool that can analyze thousands of proteins in clinical samples in one single experiment. By identifying differences in protein expression profiles, biomarkers for various diseases can hopefully be found. To date, antibody microarrays with an overall foot print of < 1 cm2, based on 18x103 µm2 (⌀ ~150 μm) sized spots at a density of ≤ 2,000 spots/cm2, have mainly been produced. Considering the size and complexity of the human proteome, this microarray design will not be able to harbor the number of antibodies required to perform global proteomics, demonstrating the need for novel, miniaturized high-density array designs. Increasing the number of antibodies on the arrays will generate a logistical problem when it comes to purification and immobilization of antibodies, which requires new technical solutions. The aim of this thesis was to develop new methods to produce the next generation of antibody arrays towards global proteome analysis. The first part of this thesis has involved design of miniaturized antibody arrays. The second part has been focused on developing antibodies with a photoreactive property, with the long-term goal to facilitate the logistics when it comes to purification and immobilization of antibodies. In conclusion, new methods to fabricate miniaturized antibody arrays, focusing on both miniaturization and antibody immobilization, for large-scale protein analysis of clinical samples have been designed. In the future, this could lead to better discovery tools for identifying biomarkers for improved diagnosis, prognosis and choice of treatments for various diseases.
|
|
