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Wester, Kenneth
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Publications (10 of 38) Show all publications
Carlsson, J., Wester, K., De La Torre, M., Malmström, P.-U. & Gardmark, T. (2015). EGFR-expression in primary urinary bladder cancer and corresponding metastases and the relation to HER2-expression. On the possibility to target these receptors with radionuclides. Radiology and Oncology, 49(1), 50-58
Open this publication in new window or tab >>EGFR-expression in primary urinary bladder cancer and corresponding metastases and the relation to HER2-expression. On the possibility to target these receptors with radionuclides
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2015 (English)In: Radiology and Oncology, ISSN 1318-2099, E-ISSN 1581-3207, Vol. 49, no 1, p. 50-58Article in journal (Refereed) Published
Abstract [en]

Background. There is limited effect of tyrosine kinase inhibitors or "naked" antibodies binding EGFR or HER2 for therapy of metastasized urinary bladder canter and these methods are therefore not routinely used. Targeting radionuclides to the extracellular domain of the receptors is potentially a better possibility. Methods. EGFR- and HER2-expression was analyzed for primary tumors and corresponding metastases from 72 patients using immunohistochemistry and the internationally recommended HercepTest. Intracellular mutations were not analyzed since only the receptors were considered as targets and intracellular abnormalities should have minor effect on radiation dose. Results. EGFR was positive in 71% of the primary tumors and 69% of corresponding metastases. Local and distant metastases were EGFR-positive in 75% and 66% of the cases, respectively. The expression frequency of HER2 in related lesions was slightly higher (data from previous study). The EGFR-positive tumors expressed EGFR in metastases in 86% of the cases. The co-expression of EGFR and HER2 was 57% for tumors and 53% for metastases. Only 3% and 10% of the lesions were negative for both receptors in tumors and metastases, respectively. Thus, targeting these receptors with radionuclides might be applied for most patients. Conclusions. At least one of the EGFR- or HER2-receptors was present in most cases and co-expressed in more than half the cases. It is therefore interesting to deliver radionuclides for whole-body receptor-analysis, dosimetry and therapy. This can hopefully compensate for resistance to other therapies and more patients can hopefully be treated with curative instead of palliative intention.

Keyword
EGFR, HER2, radionuclides, resistance, urinary bladder cancer metastases
National Category
Radiology, Nuclear Medicine and Medical Imaging Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-251515 (URN)10.2478/raon-2014-0015 (DOI)000350712100008 ()25810701 (PubMedID)
Available from: 2015-04-23 Created: 2015-04-20 Last updated: 2017-12-04Bibliographically approved
Algenas, C., Agaton, C., Fagerberg, L., Asplund, A., Bjorling, L., Bjorling, E., . . . Hober, S. (2014). Antibody performance in western blot applications is context-dependent. Biotechnology Journal, 9(3), 435-445
Open this publication in new window or tab >>Antibody performance in western blot applications is context-dependent
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2014 (English)In: Biotechnology Journal, ISSN 1860-6768, E-ISSN 1860-7314, Vol. 9, no 3, p. 435-445Article in journal (Refereed) Published
Abstract [en]

An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data.

Keyword
Immunohistochemistry, Monoclonal antibodies, Polyclonal antibodies, Validation, Western blot
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-222744 (URN)10.1002/biot.201300341 (DOI)000332339100014 ()
Available from: 2014-04-14 Created: 2014-04-14 Last updated: 2017-12-05Bibliographically approved
Gedda, L., Björkelund, H., Lebel, L., Asplund, A., Dubois, L., Wester, K., . . . Andersson, K. (2013). Evaluation of Real-Time Immunohistochemistry and Interaction Map as an Alternative Objective Assessment of HER2 Expression in Human Breast Cancer Tissue. Applied immunohistochemistry & molecular morphology (Print), 21(6), 497-505
Open this publication in new window or tab >>Evaluation of Real-Time Immunohistochemistry and Interaction Map as an Alternative Objective Assessment of HER2 Expression in Human Breast Cancer Tissue
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2013 (English)In: Applied immunohistochemistry & molecular morphology (Print), ISSN 1541-2016, E-ISSN 1533-4058, Vol. 21, no 6, p. 497-505Article in journal (Refereed) Published
Abstract [en]

Immunohistochemical study (IHC) is a critical tool in the clinical diagnosis of breast cancer. One common assessment is the expression level of the HER2 receptor in breast cancer tissue samples with the aim of stratifying patients for applicability of the therapeutic antibody Herceptin. In this study, we aimed to investigate whether a novel assay, real-time IHC combined with Interaction Map analysis, offers the possibility of objective assessment of HER2 expression. Interaction Map presents real-time interaction data as a collection of peaks on a surface, and it was performed on 20 patient tissue samples previously scored for HER2 expression. The result shows that the relative weight of the peaks in the maps contains novel information that could discriminate between high and low HER2 expression in an operator-independent manner (P<0.001). We conclude that the real-time IHC assay has a promising potential to complement conventional IHC and may improve the precision in the future clinical diagnostics of breast cancer.

Keyword
antibody, HER2, immunohistochemistry, IHC, kinetics, real time, tissue
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-212827 (URN)10.1097/PAI.0b013e318281162d (DOI)000327212100004 ()
Available from: 2013-12-18 Created: 2013-12-16 Last updated: 2017-12-06Bibliographically approved
Lindén, M., Segersten, U., Runeson, M., Wester, K., Busch, C., Pettersson, U., . . . Malmström, P.-U. (2013). Tumour expression of bladder cancer-associated urinary proteins. BJU International, 112(3), 407-415
Open this publication in new window or tab >>Tumour expression of bladder cancer-associated urinary proteins
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2013 (English)In: BJU International, ISSN 1464-4096, E-ISSN 1464-410X, Vol. 112, no 3, p. 407-415Article in journal (Refereed) Published
Abstract [en]

WHAT'S KNOWN ON THE SUBJECT? AND WHAT DOES THE STUDY ADD?:

  • The current basis for diagnosis and prognosis in urinary bladder cancer is based on the pathologists' assessment of a biopsy of the tumour. Urinary biomarkers are preferable as they can be non-invasively sampled. Urinary cytology is the only test with widespread use but is hampered by poor reproducibility and low sensitivity.
  • By studying the protein expression in bladder tumour tissue samples of proteins previously found in elevated levels in the urine of patients with bladder cancer, we have been able to show that these proteins originate from the tumour. The immunoreactivity of three of the investigated proteins increased with higher stage. Also a serine peptidase inhibitor was found to be predictive of progression from non-muscle-invasive to muscle-invasive tumours.

OBJECTIVES:

  • To analyse the expression of five bladder cancer-associated urinary proteins and investigate if expression is related to the malignant phenotype of the tumour.
  • To explore the possible prognostic value of these proteins.

PATIENTS AND METHODS:

  • Urine samples, 16 from patients with bladder cancer and 26 from controls, were used in Western Blotting experiments.
  • Tissue microarrays with bladder tissue from 344 patients diagnosed with bladder cancer between 1984 and 2005 was used in immunohistochemistry experiments.
  • The proteins apolipoprotein E (APOE), fibrinogen β chain precursor (FGB), leucine-rich α2-glycoprotein (LRG1), polymerase (RNA) I polypeptide E (POLR1E), α1-antitrypsin (SERPINA1) and topoisomerase 2A (TOP2A) were probed with antibodies validated by the Human Protein Atlas.

RESULTS:

  • Increased expressions of APOE, FGB and POLR1E were correlated with increased tumour stage (P < 0.001).
  • Expression of SERPINA1 in Ta and T1 tumours was found to increase the risk of tumour progression (hazard ratio 2.57, 95% confidence interval 1.13-5.87; P = 0.025)

CONCLUSIONS:

  • All proteins previously detected in urine from patients with bladder cancer were also expressed in bladder cancer tissue.
  • The expression of APOE, FGB and POLR1E increased with stage and they are potential diagnostic markers.
  • SERPINA1 was identified as a prognostic marker candidate.
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-197160 (URN)10.1111/j.1464-410X.2012.11653.x (DOI)000321429800024 ()23470167 (PubMedID)
Note

De två sista författarna delar sistaförfattarskapet.

Available from: 2013-03-18 Created: 2013-03-18 Last updated: 2017-12-06Bibliographically approved
Sandström, K., Haylock, A.-K., Spiegelberg, D., Qvarnström, F., Wester, K. & Nestor, M. (2012). A novel CD44v6 targeting antibody fragment with improved tumor-to-blood ratio. International Journal of Oncology, 40(5), 1525-1532
Open this publication in new window or tab >>A novel CD44v6 targeting antibody fragment with improved tumor-to-blood ratio
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2012 (English)In: International Journal of Oncology, ISSN 1019-6439, Vol. 40, no 5, p. 1525-1532Article in journal (Refereed) Published
Abstract [en]

The chimeric monoclonal antibody U36 (cMAb U36) recognizes the CD44v6 antigen. Its potential as a radioimmunotargeting agent, as well as its safety, has been shown in previous studies in head and neck cancer patients. However, intact MAbs have long circulation time in the blood and tumor targeting may also be hampered due to the slow and incomplete diffusion into solid tumors. In comparison, smaller monovalent Fab' and divalent F(ab')2 fragments are expected to exhibit shorter circulating half-lives, better tumor penetration and are thus more likely to yield better imaging results. In this study, novel F(ab')2 and Fab' fragments from cMAb U36 were radiolabeled with 125I and the characteristics of the conjugates in vitro were examined. The biodistribution of the conjugates were then evaluated in nude mice bearing CD44v6-expressing xenograft tumors. Furthermore, the penetration depth and distribution in tumor tissue was assessed by autoradiography in selected tumor samples. The in vitro experiments showed that the conjugates were stable and had intact affinity to CD44v6. The biodistribution study demonstrated superior tumor-to-blood ratio for the novel cMAb U36 fragment 125I-F(ab')2 compared with both the intact MAb and the monovalent fragment form. Autoradiography also revealed better tumor penetration for 125I-F(ab')2. This study demonstrates that the use of antibody fragments may improve radioimmunotargeting and possibly improve the management of head and neck malignancies.

National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Otorhinolaryngology
Research subject
Oto-Rhino-Laryngology
Identifiers
urn:nbn:se:uu:diva-170945 (URN)10.3892/ijo.2012.1352 (DOI)000302273400023 ()22307465 (PubMedID)
Available from: 2012-03-14 Created: 2012-03-14 Last updated: 2017-12-07Bibliographically approved
Fagelskiold, A. J., Kannisto, K., Bostrom, A., Hadrovic, B., Farre, C., Eweida, M., . . . Islam, M. S. S. (2012). Insulin-secreting INS-1E cells express functional TRPV1 channels. Islets, 4(1), 56-63
Open this publication in new window or tab >>Insulin-secreting INS-1E cells express functional TRPV1 channels
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2012 (English)In: Islets, ISSN 1938-2014, Vol. 4, no 1, p. 56-63Article in journal (Refereed) Published
Abstract [en]

We have studied whether functional TRPV1 channels exist in the INS-1E cells, a cell type used as a model for beta-cells, and in primary beta-cells from rat and human. The effects of the TRPV1 agonists capsaicin and AM404 on the intracellular free Ca2+ concentration ([Ca2+]i) in the INS-1E cells were studied by fura-2based microfluorometry. Capsaicin increased [Ca2+] i in a concentration-dependent manner, and the [Ca2+] i increase was dependent on extracellular Ca2+. AM404 also increased [Ca2+] i in the INS-1E cells. Capsazepine, a specific antagonist of TRPV1, completely blocked the capsaicin- and AM404-induced [Ca2+] i increases. Capsaicin did not increase [Ca2+] i in the primary beta-cells from rat and human. Whole cell patch clamp configuration was used to record currents across the plasma membrane in the INS-1E cells. Capsaicin elicited inward currents that were inhibited by capsazepine. Western blot analysis detected TRPV1 proteins in the INS-1E cells and the human islets. Immunohistochemistry was used to study the expression of TRPV1, but no TRPV1 protein immunoreactivity was detected in the human islet cells and the human insulinoma cells. We conclude that the INS-1E cells, but not the primary beta-cells, express functional TRPV1 channels.

Keyword
cell signaling, Ca2+, INS-1E cells, TRPV1, capsaicin, and AM404
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-175870 (URN)10.4161/isl.18915 (DOI)000303980000008 ()
Available from: 2012-06-13 Created: 2012-06-13 Last updated: 2012-06-13Bibliographically approved
Lindén, M., Lind, S. B., Mayrhofer, C., Segersten, U., Wester, K., Lyutvinskiy, Y., . . . Pettersson, U. (2012). Proteomic analysis of urinary biomarker candidates for nonmuscle invasive bladder cancer. Proteomics, 12(1), 135-144
Open this publication in new window or tab >>Proteomic analysis of urinary biomarker candidates for nonmuscle invasive bladder cancer
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2012 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 12, no 1, p. 135-144Article in journal (Refereed) Published
Abstract [en]

Nonmuscle invasive tumors of the bladder often recur and thereby bladder cancer patients need regular re-examinations which are invasive, unpleasant, and expensive. A noninvasive and less expensive method, e.g. a urine dipstick test, for monitoring recurrence would thus be advantageous. In this study, the complementary techniques mass spectrometry (MS) and Western blotting (WB)/dot blot (DB) were used to screen the urine samples from bladder cancer patients. High resolving MS was used to analyze and quantify the urinary proteome and 29 proteins had a significantly higher abundance (p<0.05) in bladder cancer samples compared with control urine samples. The increased abundance found in urine from bladder cancer patients compared with controls was confirmed with Western blot for four selected proteins; fibrinogen β chain precursor, apolipoprotein E, α-1-antitrypsin, and leucine-rich α-2-glycoprotein 1. Dot blot analysis of an independent urine sample set pointed out fibrinogen β chain and α-1-antitrypsin as most interesting biomarkers having sensitivity and specificity values in the range of 66-85%. Exploring the Human Protein Atlas (HPA) also revealed that bladder cancer tumors are the likely source of these proteins. They have the potential of being useful in diagnosis, monitoring of recurrence and thus may improve the treatment of bladder tumors, especially nonmuscle invasive tumors.

National Category
Urology and Nephrology Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-163407 (URN)10.1002/pmic.201000810 (DOI)000298841000016 ()22065568 (PubMedID)
Available from: 2011-12-12 Created: 2011-12-12 Last updated: 2017-12-08Bibliographically approved
Malmberg, J., Sandström, M., Wester, K., Tolmachev, V. & Orlova, A. (2011). Comparative biodistribution of imaging agents for in vivo molecular profiling of disseminated prostate cancer in mice bearing prostate cancer xenografts: focus on (111)In- and (125)I-labeled anti-HER2 humanized monoclonal trastuzumab and ABY-025 Affibody. Nuclear Medicine and Biology, 38(8), 1093-1102
Open this publication in new window or tab >>Comparative biodistribution of imaging agents for in vivo molecular profiling of disseminated prostate cancer in mice bearing prostate cancer xenografts: focus on (111)In- and (125)I-labeled anti-HER2 humanized monoclonal trastuzumab and ABY-025 Affibody
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2011 (English)In: Nuclear Medicine and Biology, ISSN 0969-8051, E-ISSN 1872-9614, Vol. 38, no 8, p. 1093-1102Article in journal (Refereed) Published
Abstract [en]

Introduction: Human epidermal growth factor receptor type 2 (HER2) overexpression supports proliferation of androgen-independent prostate cancer (PC). Radionuclide molecular imaging of HER2 expression in disseminated PC would aid in the selection of patients who are likely responders to HER2 targeting therapy. In this study, we evaluated whether ABY-025 Affibody molecule, a small (similar to 7-kDa) HER2-binding scaffold protein, produces superior tumor-to-nontumor ratios compared with those obtained through the use of radiolabeled humanized anti-HER2 antibody, trastuzumab. The influence of (111)In vs. (125)I radiolabel was evaluated for both tracers.

Methods: ABY-025 was labeled with (111)In using 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid chelator, site-specifically coupled to the C-terminus via the maleimido derivative. Trastuzumab was labeled with (111)In using a CHX-A" diethylene triamine pentaacetic acid (DTPA) chelator. An indirect radioiodination with [(125)I]-N-succinimidyl-para-iodobenzoate was used for both targeting proteins. Biodistribution of all labeled targeting proteins was evaluated in mice bearing DU-145 PC xenografts.

Results: The use of residualizing (111)In-label facilitated better tumor uptake and better tumor-to-nontumor ratios for both targeting agents. [(111)In]-ABY-025 provided tumor uptake of 7.1 +/- 0.8% injected dose per gram of tissue (% ID/g) and tumor-to-blood ratio of 47 +/- 13 already at 6 h postinjection. The maximum tumor-to-nontumor ratios with [(111)In]-CHX-DTPA-trastuzumab were achieved at 72 h postinjection, whereas tumor uptake was 11 +/- 4% ID/g and tumor-to-blood ratio was 18 +/- 7. The biodistribution data were confirmed with gamma-camera imaging.

Conclusions: Radiolabeled ABY-025 Affibody molecule provides higher contrast in imaging of HER2-expressing PC xenografts than radiolabeled trastuzumab. Residualizing radiometal label for ABY-025 provides better contrast in imaging of HER2-expressing PC xenografts than nonresidualizing radiohalogen.

Keyword
HER2, Prostate cancer, Trastuzumab, Affibody molecule, Biodistribution, Radiolabeling
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-166866 (URN)10.1016/j.nucmedbio.2011.04.005 (DOI)000298070400003 ()22137850 (PubMedID)
Available from: 2012-01-16 Created: 2012-01-16 Last updated: 2017-12-08Bibliographically approved
Segersten, M. U., Edlund, E. K., Micke, P., de la Torre, M., Hamberg, H., Edvinsson, Å. E. L., . . . Wester, H. K. (2009). A novel strategy based on histological protein profiling in-silico for identifying potential biomarkers in urinary bladder cancer. BJU International, 104(11), 1780-1785
Open this publication in new window or tab >>A novel strategy based on histological protein profiling in-silico for identifying potential biomarkers in urinary bladder cancer
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2009 (English)In: BJU International, ISSN 1464-4096, E-ISSN 1464-410X, Vol. 104, no 11, p. 1780-1785Article in journal (Refereed) Published
Abstract [en]

OBJECTIVE: To screen a publicly available immunohistochemistry (IHC) based web-atlas, to identify key proteins in bladder cancer that might serve as potential biomarkers. MATERIALS AND METHODS: The first version of the Human Protein Atlas (HPA 1.0), with 660 proteins, was visually examined to identify proteins with a variable staining pattern among the 12 tissue samples representing bladder cancer. None or limited previous characterization in bladder cancer, as well as a supportive Western blot, were also required. The selected proteins were then evaluated in an independent set of patient samples (106 tumour samples of differing stage and grade) represented in a tissue microarray (TMAi). The IHC expression of the identified proteins in the TMAi was scored and related to tumour stage and grade. RESULTS: The expression profiles of the 13 proteins selected from the web-atlas were confirmed in the TMAi. Expression patterns for seven proteins were significantly altered (P < 0.05) with higher stage and/or grade. Three of those (CN130, DSG3, PHF6) lack characterization in bladder cancer, whereas the remaining four proteins have previously been suggested as key proteins/potential biomarkers in cancer, some of them also in bladder cancer. CONCLUSION: New candidate proteins for urinary bladder cancer were identified through screening of the publicly available HPA 1.0. Although further evaluation is necessary, this strategy is promising in the search for new biomarkers, with potential to improve the management of patients with this disease.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-112911 (URN)10.1111/j.1464-410X.2009.08674.x (DOI)000271627100044 ()19522865 (PubMedID)
Available from: 2010-01-22 Created: 2010-01-22 Last updated: 2018-02-01Bibliographically approved
Larsson, K., Eriksson, C., Schwenk, J. M., Berglund, L., Wester, K., Uhlén, M., . . . Wernérus, H. (2009). Characterization of PrEST-based antibodies towards human Cytokeratin-17. JIM - Journal of Immunological Methods, 342(1-2), 20-32
Open this publication in new window or tab >>Characterization of PrEST-based antibodies towards human Cytokeratin-17
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2009 (English)In: JIM - Journal of Immunological Methods, ISSN 0022-1759, E-ISSN 1872-7905, Vol. 342, no 1-2, p. 20-32Article in journal (Refereed) Published
Abstract [en]

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 specific antibodies. In this study, two non-overlapping parts of human Cytokeratin-17, a protein belonging to the intermediate filament family of highly sequence-related proteins, were selected as a model system to study the specificity and cross reactivity of antibodies generated towards such a target. These recombinantly produced Protein Epitope Signature Tags (PrESTs) were immunized in five rabbits each and the batch-to-batch variations in the obtained immune responses were studied by mapping of linear epitopes using synthetic overlapping peptides. The obtained results showed a similar but not identical immune response in the respective antibody groups with a limited number of epitopes being identified. Immunohistochemical analysis of the affinity purified monospecific antibodies on tissue micro arrays resulted in a general recognition of human cytokeratins for all analyzed binders whereas antibodies identified as binding to the most unique parts of the PrESTs showed the most Cytokeratin-17 like staining. The data presented here support the strategy to use sequence identity scores as the main criteria for antigen selection but also indicate the possibility to instead produce a single antibody recognizing a defined group of proteins when the intended targets overall sequence identity score is too high. This type of group-specific antibodies would be an important tool for antibody-based projects aiming for a complete coverage of the human proteome.

Keyword
Cytokeratin-17, PrEST, Epitope mapping, Antibody
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-103930 (URN)10.1016/j.jim.2008.11.013 (DOI)000264685300003 ()19108777 (PubMedID)
Available from: 2009-05-26 Created: 2009-05-26 Last updated: 2017-12-13Bibliographically approved
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