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Johansson, K. & Mohlin, C. (2024). Qualitative evaluations of reactive microglial heterogeneity in cultured porcine retina. Histology and Histopathology, 39(12), 1611-1620
Open this publication in new window or tab >>Qualitative evaluations of reactive microglial heterogeneity in cultured porcine retina
2024 (English)In: Histology and Histopathology, ISSN 0213-3911, E-ISSN 1699-5848, Vol. 39, no 12, p. 1611-1620Article in journal (Refereed) Published
Abstract [en]

A late stage of several retinal disorders is retinal detachment, a complication that results in rapid photoreceptor degeneration and synaptic damage. The porcine retina is a favorable in vitro model for studies of the degenerative processes that follow retinal detachment. Photoreceptor degeneration and synaptic injuries develop rapidly in the cultured porcine retina and correlate with resident microglial cell transition into a reactive phenotype. In this in vitro study, we used retinas cultured for five days and analyzed reactive CD11b and Iba1 immunoreactive microglia that localized close to/within the synaptic outer plexiform layer (OPL) and in the outer nuclear layer (ONL). A subpopulation of the CD11b and Iba1immunoreactive microglia also expressed CD68 immunoreactivity on lysosomal membranes or as a diffuse cytoplasmic stain. Some CD68 immunoreactive microglia were juxtaposed to L/M-opsin immunoreactive cone photoreceptors in the ONL. CD11b and Iba immunoelectron microscopy further suggests the presence of a dark microglial phenotype in the degenerating cultured porcine retina. For immunoelectron microscopy, nickel-enhanced diaminobenzidine (DAB) staining resulted in clearly distinguished reaction products in the cytosol of dark microglia.

Keywords
Retina, Microglia, CD11b, Iba1, CD68, Photoreceptor, Immunoelectron microscopy
National Category
Ophthalmology
Research subject
Chemistry, Biochemistry
Identifiers
urn:nbn:se:lnu:diva-134357 (URN)10.14670/HH-18-772 (DOI)001381254900006 ()38860562 (PubMedID)2-s2.0-85209796028 (Scopus ID)
Available from: 2025-01-08 Created: 2025-01-08 Last updated: 2025-03-20Bibliographically approved
Andersson, L., Sjöström, D. J., Quach, H. Q., Hägerström, K., Hurler, L., Kajdacsi, E., . . . Nilsson, P. H. (2024). Storage of Transfusion Platelet Concentrates is Associated with Complement Activation and Reduced Ability of Platelets to Respond to Protease-Activated Receptor-1 and Thromboxane A2 Receptor. International Journal of Molecular Sciences, 25(2), Article ID 1091.
Open this publication in new window or tab >>Storage of Transfusion Platelet Concentrates is Associated with Complement Activation and Reduced Ability of Platelets to Respond to Protease-Activated Receptor-1 and Thromboxane A2 Receptor
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2024 (English)In: International Journal of Molecular Sciences, ISSN 1661-6596, E-ISSN 1422-0067, Vol. 25, no 2, article id 1091Article in journal (Refereed) Published
Abstract [en]

Platelet activation and the complement system are mutually dependent. Here, we investigated the effects of storage time on complement activation and platelet function in routinely produced platelet concentrates. The platelet concentrates (n = 10) were stored at 22 degrees C for seven days and assessed daily for complement and platelet activation markers. Additionally, platelet function was analyzed in terms of their responsiveness to protease-activated receptor-1 (PAR-1) and thromboxane A2 receptor (TXA(2)R) activation and their capacity to adhere to collagen. Complement activation increased over the storage period for all analyzed markers, including the C1rs/C1-INH complex (fold change (FC) = 1.9; p < 0.001), MASP-1/C1-INH complex (FC = 2.0; p < 0.001), C4c (FC = 1.8, p < 0.001), C3bc (FC = 4.0; p < 0.01), and soluble C5b-9 (FC = 1.7, p < 0.001). Furthermore, the levels of soluble platelet activation markers increased in the concentrates over the seven-day period, including neutrophil-activating peptide-2 (FC = 2.5; p < 0.0001), transforming growth factor beta 1 (FC = 1.9; p < 0.001) and platelet factor 4 (FC = 2.1; p < 0.0001). The ability of platelets to respond to activation, as measured by surface expression of CD62P and CD63, decreased by 19% and 24% (p < 0.05) for PAR-1 and 69-72% (p < 0.05) for TXA(2)R activation, respectively, on Day 7 compared to Day 1. The extent of platelet binding to collagen was not significantly impaired during storage. In conclusion, we demonstrated that complement activation increased during the storage of platelets, and this correlated with increased platelet activation and a reduced ability of the platelets to respond to, primarily, TXA(2)R activation.

Place, publisher, year, edition, pages
MDPI, 2024
Keywords
platelet storage, platelet storage lesion, complement activation, platelet function, hemostasis
National Category
Immunology Cell Biology
Research subject
Biomedical Sciences, Immunology; Natural Science, Cell and Organism Biology
Identifiers
urn:nbn:se:lnu:diva-127877 (URN)10.3390/ijms25021091 (DOI)001152950800001 ()38256162 (PubMedID)2-s2.0-85183338259 (Scopus ID)
Available from: 2024-02-20 Created: 2024-02-20 Last updated: 2025-02-04Bibliographically approved
Baskaran, K., Nilsson, I., Ringbäck, K., Ternehäll, M., Svanfeldt, C., Melin, J., . . . Macedo, A. F. (2024). Swedish version of the Massof activity inventory to measure vision-related activity difficulties among patients with nAMD. In: Acta Ophthalmol, vol 102, issue S279: Special Issue:Abstracts from the 2023 European Association for Vision and Eye Research Festival, 26‐28 October 2023, Valencia: . Paper presented at 2023 European Association for Vision and Eye Research Festival, 26‐28 October 2023, Valencia. John Wiley & Sons, 102(S279)
Open this publication in new window or tab >>Swedish version of the Massof activity inventory to measure vision-related activity difficulties among patients with nAMD
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2024 (English)In: Acta Ophthalmol, vol 102, issue S279: Special Issue:Abstracts from the 2023 European Association for Vision and Eye Research Festival, 26‐28 October 2023, Valencia, John Wiley & Sons, 2024, Vol. 102, no S279Conference paper, Oral presentation with published abstract (Refereed)
Abstract [en]

Aims/Purpose: The aim of this study was to assess vision-related activity difficulties among patients with neovascular AMD using a Swedish version of the mass of activity inventory (MAI). Methods: Participants were patients diagnosed with neovascular AMD receiving treatment for the disease in a hospital in southeast Sweden. Participants completed the Swedish version of the MAI questionnaire. MAI can be used to measure the overall visual ability and visual ability in 4 functional domains: reading, mobility, visual motor function and visual information processing. Best corrected distance and near visual acuity (VA) were also measured. Results: Among the 196 participants (mean age=78.5 years, SD=7.67, 66% female) the median VA in the better seeing eye was 0.18 logMAR (IQR=?0.34), and in the worse eye was 0.54 logMAR (IQR=0.98). The median visual ability for all participants was 1.92 logits (IQR=2.69). There was a significant negative correlation between distance VA in the better eye and visual ability (rho=0.4025, p<0.01). Using ROC curves, we tested the capacity of the MAI to detect cases of any vision impairment (VA worse than 0.3 logMAR in the better seeing eye), the area under the curve (AUC) was 0.717 (95% CI=0.643 - 0.791 p<0.001). When we tested for detection of moderate vision impairment (VA worse than 0.5 logMAR in the better seeing eye) the AUC was 0.738 (95% CI=0.648 - 0.829 p?<0.001). Conclusions: The results indicate that the Swedish version of the MAI produce measures of visual ability that are consistent with clinical measures among patients with nAMD. The Swedish version of the MAI can be used as outcome measure in interventions for people with nAMD.

References

1. Macedo, A.F. et al. Predictors of problems reported on the EQ-5D-3L dimensions among people with impaired vision in northern Portugal. Health Qual Life Outcomes 2022; 20: 132.

Place, publisher, year, edition, pages
John Wiley & Sons, 2024
National Category
Ophthalmology
Research subject
Natural Science, Optometry
Identifiers
urn:nbn:se:lnu:diva-127339 (URN)10.1111/aos.15977 (DOI)001294533400159 ()
Conference
2023 European Association for Vision and Eye Research Festival, 26‐28 October 2023, Valencia
Available from: 2024-01-31 Created: 2024-01-31 Last updated: 2025-02-18Bibliographically approved
Sjöström, D. J., Grill, B., Ambrosetti, E., Veetil, A. A., Mohlin, C., Teixeira, A. I., . . . Bjelic, S. (2023). Affinity Maturated Transferrin Receptor Apical Domain Blocks Machupo Virus Glycoprotein Binding. Journal of Molecular Biology, 435(20), Article ID 168262.
Open this publication in new window or tab >>Affinity Maturated Transferrin Receptor Apical Domain Blocks Machupo Virus Glycoprotein Binding
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2023 (English)In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 435, no 20, article id 168262Article in journal (Refereed) Published
Abstract [en]

Transferrin receptor 1 (TfR) delivers iron across cellular membranes by shuttling the ion carrier protein transferrin. This ability to deliver large protein ligands inside cells is taken advantage of by pathogens to infiltrate human cells. Notably, the receptor's outermost ectodomain, the apical domain, is used as a point of attachment for several viruses including hemorrhagic arenaviruses. To better understand interactions with the receptor it would be advantageous to probe sequence determinants in the apical domain with viral spike proteins. Here, we carried out affinity maturation of our computationally designed apical domain from human TfR to identify underlying driving forces that lead to better binding. The improved variants were confirmed by in vitro surface plasmon resonance measurements with dissociation constants obtained in the lower nanomolar range. It was found that the strong binding affinities for the optimized variants matched the strength of interactions with the native receptor. The structure of the best variant was determined experimentally indicating that the conformational change in the hairpin binding motif at the protein-protein interface plays a crucial role. The experimental methodology can be straightforwardly applied to other arenavirus or pathogens that use the apical domain. It can further be useful to probe host-virus compatibility or therapeutic strategies based on the transferrin receptor decoys.

Place, publisher, year, edition, pages
Elsevier, 2023
Keywords
transferrin receptor, yeast surface display, Rosetta, protein design
National Category
Biochemistry Molecular Biology
Research subject
Chemistry, Biochemistry
Identifiers
urn:nbn:se:lnu:diva-125437 (URN)10.1016/j.jmb.2023.168262 (DOI)001081482800001 ()37678707 (PubMedID)2-s2.0-85171429160 (Scopus ID)
Available from: 2023-11-02 Created: 2023-11-02 Last updated: 2025-03-19Bibliographically approved
Gerogianni, A., Bal, M., Mohlin, C., Woodruff, T. M., Lambris, J. D., Mollnes, T. E., . . . Nilsson, P. H. (2023). In vitro evaluation of iron oxide nanoparticle-induced thromboinflammatory response using a combined human whole blood and endothelial cell model. Frontiers in Immunology, 14, Article ID 1101387.
Open this publication in new window or tab >>In vitro evaluation of iron oxide nanoparticle-induced thromboinflammatory response using a combined human whole blood and endothelial cell model
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2023 (English)In: Frontiers in Immunology, E-ISSN 1664-3224, Vol. 14, article id 1101387Article in journal (Refereed) Published
Abstract [en]

Iron oxide nanoparticles (IONPs) are widely used in diagnostic and therapeutic settings. Upon systemic administration, however, they are rapidly recognized by components of innate immunity, which limit their therapeutic capacity and can potentially lead to adverse side effects. IONPs were previously found to induce the inflammatory response in human whole blood, including activation of the complement system and increased secretion of cytokines. Here, we investigated the thromboinflammatory response of 10-30 nm IONPs in lepirudin anticoagulated whole blood in interplay with endothelial cells and evaluated the therapeutic effect of applying complement inhibitors to limit adverse effects related to thromboinflammation. We found that IONPs induced complement activation, primarily at the C3-level, in whole blood incubated for up to four hours at 37°C with and without human microvascular endothelial cells. Furthermore, IONPs mediated a strong thromboinflammatory response, as seen by the significantly increased release of 21 of the 27 analyzed cytokines (p<0.05). IONPs also significantly increased cell-activation markers of endothelial cells [ICAM-1 (p<0.0001), P/E-selectin (p<0.05)], monocytes, and granulocytes [CD11b (p<0.001)], and platelets [CD62P (p<0.05), CD63 (p<0.05), NAP-2 (p<0.01), PF4 (p<0.05)], and showed cytotoxic effects, as seen by increased LDH (p<0.001) and heme (p<0.0001) levels. We found that inflammation and endothelial cell activation were partly complement-dependent and inhibition of complement at the level of C3 by compstatin Cp40 significantly attenuated expression of ICAM-1 (p<0.01) and selectins (p<0.05). We show that complement activation plays an important role in the IONPs-induced thromboinflammatory response and that complement inhibition is promising in improving IONPs biocompatibility.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2023
National Category
Immunology
Research subject
Biomedical Sciences, Immunology
Identifiers
urn:nbn:se:lnu:diva-118763 (URN)10.3389/fimmu.2023.1101387 (DOI)000970005700001 ()37081885 (PubMedID)2-s2.0-85153430330 (Scopus ID)
Note

Is included in the dissertation as a manuscript.

Available from: 2023-01-26 Created: 2023-01-26 Last updated: 2025-02-04Bibliographically approved
Johansson, K. & Mohlin, C. (2023). Microglia in Cultured Porcine Retina: Qualitative Immunohistochemical Analyses of Reactive Microglia in the Outer Retina. International Journal of Molecular Sciences, 24(1), Article ID 871.
Open this publication in new window or tab >>Microglia in Cultured Porcine Retina: Qualitative Immunohistochemical Analyses of Reactive Microglia in the Outer Retina
2023 (English)In: International Journal of Molecular Sciences, ISSN 1661-6596, E-ISSN 1422-0067, Vol. 24, no 1, article id 871Article in journal (Refereed) Published
Abstract [en]

A late stage of several retinal disorders is retinal detachment, a complication that results in rapid photoreceptor degeneration and synaptic damages. Experimental retinal detachment in vivo is an invasive and complicated method performed on anesthetized animals. As retinal detachment may result in visual impairment and blindness, research is of fundamental importance for understanding degenerative processes. Both morphological and ethical issues make the porcine retina a favorable organotypic model for studies of the degenerative processes that follow retinal detachment. In the cultured retina, photoreceptor degeneration and synaptic injuries develop rapidly and correlate with resident microglial cells' transition into a reactive phenotype. In this immunohistochemical study, we have begun to analyze the transition of subsets of reactive microglia which are known to localize close to the outer plexiform layer (OPL) in degenerating in vivo and in vitro retina. Biomarkers for reactive microglia included P2Ry12, CD63 and CD68 and the general microglial markers were CD11b, Iba1 and isolectin B-4 (IB4). The reactive microglia markers labeled microglia subpopulations, suggesting that protective or harmful reactive microglia may be present simultaneously in the injured retina. Our findings support the usage of porcine retina cultures for studies of photoreceptor injuries related to retinal detachment.

Place, publisher, year, edition, pages
MDPI, 2023
Keywords
retina, CD11b, Iba1, reactive microglia, synapse
National Category
Ophthalmology Neurosciences
Research subject
Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-118834 (URN)10.3390/ijms24010871 (DOI)000910450900001 ()36614320 (PubMedID)2-s2.0-85145945472 (Scopus ID)
Available from: 2023-01-30 Created: 2023-01-30 Last updated: 2023-02-21Bibliographically approved
Gerogianni, A., Dimitrov, J. D., Zarantonello, A., Poillerat, V., Chonat, S., Sandholm, K., . . . Nilsson, P. H. (2022). Heme Interferes With Complement Factor I-Dependent Regulation by Enhancing Alternative Pathway Activation. Frontiers in Immunology, 13, Article ID 901876.
Open this publication in new window or tab >>Heme Interferes With Complement Factor I-Dependent Regulation by Enhancing Alternative Pathway Activation
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2022 (English)In: Frontiers in Immunology, E-ISSN 1664-3224, Vol. 13, article id 901876Article in journal (Refereed) Published
Abstract [en]

Hemolysis, as a result of disease or exposure to biomaterials, is characterized by excess amounts of cell-free heme intravascularly and consumption of the protective heme-scavenger proteins in plasma. The liberation of heme has been linked to the activation of inflammatory systems, including the complement system, through alternative pathway activation. Here, we investigated the impact of heme on the regulatory function of the complement system. Heme dose-dependently inhibited factor I-mediated degradation of soluble and surface-bound C3b, when incubated in plasma or buffer with complement regulatory proteins. Inhibition occurred with factor H and soluble complement receptor 1 as co-factors, and the mechanism was linked to the direct heme-interaction with factor I. The heme-scavenger protein hemopexin was the main contaminant in purified factor I preparations. This led us to identify that hemopexin formed a complex with factor I in normal human plasma. These complexes were significantly reduced during acute vasoocclusive pain crisis in patients with sickle cell disease, but the complexes were normalized at their baseline outpatient clinic visit. Hemopexin exposed a protective function of factor I activity in vitro, but only when it was present before the addition of heme. In conclusion, we present a mechanistic explanation of how heme promotes uncontrolled complement alternative pathway amplification by interfering with the regulatory capacity of factor I. Reduced levels of hemopexin and hemopexin-factor I complexes during an acute hemolytic crisis is a risk factor for heme-mediated factor I inhibition.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2022
Keywords
heme, complement, factor I, co-factor activity, hemopexin, hemolysis
National Category
Immunology
Research subject
Biomedical Sciences, Immunology
Identifiers
urn:nbn:se:lnu:diva-116295 (URN)10.3389/fimmu.2022.901876 (DOI)000837093000001 ()35935964 (PubMedID)2-s2.0-85135475624 (Scopus ID)
Available from: 2022-09-16 Created: 2022-09-16 Last updated: 2024-01-17Bibliographically approved
Sjöström, D. J., Mohlin, C., Ambrosetti, E., Garforth, S. J., Teixeira, A. I. & Bjelic, S. (2022). Motif-driven protein binder design towards transferrin receptor helical domain. The FEBS Journal, 289(10), 2935-2947
Open this publication in new window or tab >>Motif-driven protein binder design towards transferrin receptor helical domain
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2022 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 289, no 10, p. 2935-2947Article in journal (Refereed) Published
Abstract [en]

Human transferrin receptor 1 (TfR) is necessary for the delivery of the iron carrier protein transferrin into cells and can be utilized for targeted delivery across cellular membranes. Binding of transferrin to the receptor is regulated by hereditary hemochromatosis protein (HFE), an iron regulatory protein that partly shares a binding site with transferrin on TfR. Here, we derived essential binding interactions from HFE and computationally grafted these into a library of small protein scaffolds. One of the designed proteins, TB08, was further optimized computationally and experimentally to identify variants with improved binding to TfR. The optimized variant, TB08 S3.1, expressed well in the E. coli expression system and had an affinity to TfR in the low micromolar range, K-d approximate to 1 mu m, as determined by surface plasmon resonance. A binding competition assay with transferrin further confirmed the interaction of the evolved variant to TfR at the shared binding surface. Additionally, the GFP-tagged evolved variant of TB08 demonstrated cellular internalization as determined by fluorescent and confocal microscopy in HeLa cells. The designed protein is small, allows for robust cargo tagging, and interacts specifically with TfR, thus making it a valuable tool for the characterization of TfR-mediated cellular transport mechanisms and for the assessment of engineering strategies for cargo delivery across cell membranes.

Place, publisher, year, edition, pages
John Wiley & Sons, 2022
Keywords
protein design, rosetta, transferrin receptor, yeast surface display
National Category
Biochemistry Molecular Biology
Research subject
Chemistry, Biochemistry
Identifiers
urn:nbn:se:lnu:diva-109632 (URN)10.1111/febs.16311 (DOI)000729487000001 ()34862739 (PubMedID)2-s2.0-85121011677 (Scopus ID)2021 (Local ID)2021 (Archive number)2021 (OAI)
Available from: 2022-01-20 Created: 2022-01-20 Last updated: 2025-02-20Bibliographically approved
Islam, R., Islam, M. M., Nilsson, P. H., Mohlin, C., Hagen, K. T., Paschalis, E. I., . . . Mollnes, T. E. (2021). Combined blockade of complement C5 and TLR co-receptor CD14 synergistically inhibits pig-to-human corneal xenograft induced innate inflammatory responses. Acta Biomaterialia, 127, 169-179
Open this publication in new window or tab >>Combined blockade of complement C5 and TLR co-receptor CD14 synergistically inhibits pig-to-human corneal xenograft induced innate inflammatory responses
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2021 (English)In: Acta Biomaterialia, ISSN 1742-7061, E-ISSN 1878-7568, Vol. 127, p. 169-179Article in journal (Refereed) Published
Abstract [en]

Inadequate supplies of donor corneas have evoked an escalating interest in corneal xenotransplantation. However, innate immune responses contribute significantly to the mechanism of xenograft rejection. We hypothesized that complement component C5 and TLR co-receptor CD14 inhibition would inhibit porcine cornea induced innate immune responses. Therefore, we measured cytokine release in human blood, induced by three forms of corneal xenografts with or without inhibitors. Native porcine cornea (NPC) induced interleukins (IL-1 beta, IL-2, IL-6, IL-8, IL-1ra), chemokines (MCP-1, MIP-1 alpha, MIP-1 beta) and other cytokines (TNF, G-CSF, INF-gamma, FGF-basic). Decellularized (DPC) and gamma-irradiated cornea (g-DPC) elevated the release of those cytokines. C5-blockade by eculizumab inhibited all the cytokines except G-CSF when induced by NPC. However, C5-blockade failed to reduce DPC and g-DPC induced cytokines. Blockade of CD14 inhibited DPC-induced cytokines except for IL-8, MCP-1, MIP-1 alpha, and G-CSF, while it inhibited all of them when induced by g-DPC. Combined blockade of C5 and CD14 inhibited the maximum number of cytokines regardless of the xenograft type. Finally, by using the TLR4 specific inhibitor Eritoran, we showed that TLR4 activation was the basis for the CD14 effect. Thus, blockade of C5, when combined with TLR4 inhibition, may have therapeutic potential in pig-to-human corneal xenotransplantation. Statement of significance Bio-engineered corneal xenografts are on the verge of becoming a viable alternative to allogenic human donor-cornea, but the host's innate immune response is still a critical barrier for graft acceptance. By overruling this barrier, limited graft availability would no longer be an issue for treating corneal diseases. We showed that the xenograft induced inflammation is initiated by the complement system and toll-like receptor activation. Intriguingly, the inflammatory response was efficiently blocked by simultaneously targeting bottleneck molecules in the complement system (C5) and the TLR co-receptor CD14 with pharmaceutical inhibitors. We postulate that a combination of C5 and CD14 inhibition could have a great therapeutic potential to overcome the immunologic barrier in pig-to-human corneal xenotransplantation. (C) 2021 The Authors. Published by Elsevier Ltd on behalf of Acta Materialia Inc.

Place, publisher, year, edition, pages
Elsevier, 2021
Keywords
Xenotransplantation, Cornea, Decellularization, Complement system, Toll-like receptor, Cytokine, Regenerative medicine, Biomaterial
National Category
Immunology in the medical area Biomaterials Science
Research subject
Biomedical Sciences, Immunology
Identifiers
urn:nbn:se:lnu:diva-105799 (URN)10.1016/j.actbio.2021.03.047 (DOI)000653397700011 ()33785451 (PubMedID)2-s2.0-85104291155 (Scopus ID)2021 (Local ID)2021 (Archive number)2021 (OAI)
Available from: 2021-07-08 Created: 2021-07-08 Last updated: 2023-01-24Bibliographically approved
Sandholm, K., Persson, B., Abdalla, S., Mohlin, C., Nilsson, B. & Nilsson Ekdahl, K. (2021). Quantification of Complement Proteins with Special Reference to C1q: Multiplex Versus ELISA Versus Rocket Immunoelectrophoresis Versus Nephelometry. In: Lubka T. Roumenina (Ed.), The Complement System: Methods in Molecular Biology (pp. 33-41). Humana Press, 2227
Open this publication in new window or tab >>Quantification of Complement Proteins with Special Reference to C1q: Multiplex Versus ELISA Versus Rocket Immunoelectrophoresis Versus Nephelometry
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2021 (English)In: The Complement System: Methods in Molecular Biology / [ed] Lubka T. Roumenina, Humana Press, 2021, Vol. 2227, p. 33-41Chapter in book (Refereed)
Abstract [en]

Accurate determination of complement component C1q is hampered by the fact that C1q is an immune complex binding protein. Consequently, immunochemical techniques which rely on immune complex formation in fluid phase such as nephelometry and turbidimetry tend to give results which differ from those obtained by, for example, ELISA and other solid phase-based assays. In this chapter, we discuss the pros and cons of different techniques for the quantification of C1q and present a comprehensive protocol for a newly developed magnetic bead-based sandwich immunoassay which has replaced nephelometry in our complement diagnostic laboratory at the University Hospital in Uppsala.

Place, publisher, year, edition, pages
Humana Press, 2021
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 2227
Keywords
Humans, Complement System Proteins, ELISA, Enzyme-Linked Immunosorbent Assay, Blood Protein Electrophoresis, C1q, Cerebrospinal fluid (CSF), Complement C1q, Diagnostic Tests, Routine, Immunoassays, Immunoelectrophoresis, Immunomagnetic Separation, Multiplex, Nephelometry, Nephelometry and Turbidimetry, Plasma, Rocket immunoelectrophoresis (RIE)
National Category
Immunology in the medical area
Research subject
Biomedical Sciences, Immunology; Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-112011 (URN)10.1007/978-1-0716-1016-9_3 (DOI)000679413000004 ()2-s2.0-85104284726 (Scopus ID)9781071610152 (ISBN)9781071610169 (ISBN)9781071610183 (ISBN)
Funder
Swedish Research Council, 2016-2075-5.1Swedish Research Council, 2016-04519Medical Research Council of Southeast Sweden (FORSS)
Note

This work was supported by grant 2016-2075-5.1 and 2016-04519 from the Swedish Research Council (VR), the Medical Research Council of Southeast Sweden (FORSS), and by faculty grants from the Linnaeus university. The study was also supported by grants for development and validation of complement assays provided by the University Hospital in Uppsala, Sweden.

Available from: 2022-05-03 Created: 2022-05-03 Last updated: 2022-11-09Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-9301-1977

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