Design of recombinant antibody microarrays for membrane protein profiling of cell lysates and tissue extracts

Dexlin-Mellby, Linda; Sandström, Anna; Antberg, Linn; Gunnarsson, Jenny; Hansson, Stefan R.; Borrebaeck, Carl; Wingren, Christer

doi:10.1002/pmic.200900808

Cited by 19 publications

(16 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The homogenized samples were weighted and transferred to a collection tube and stored at -80°C until use. Extraction of proteins from homogenized tissue followed previously described protocol [31, 32]. Briefly, 10 μl of Extraction Buffer (100μg/ml Soybean trypsin inhibitor, 350 μg/ml PMSF, 0.01% (w/v) BSA and 2% (w/v) Saponin) was added per mg of sample and incubated on a rocking table at 4°C o/n.…”

Section: Methodsmentioning

confidence: 99%

Tumor tissue protein signatures reflect histological grade of breast cancer

et al. 2017

Self Cite

View full text Add to dashboard Cite

Histological grade is one of the most commonly used prognostic factors for patients diagnosed with breast cancer. However, conventional grading has proven technically challenging, and up to 60% of the tumors are classified as histological grade 2, which represents a heterogeneous cohort less informative for clinical decision making. In an attempt to study and extend the molecular puzzle of histologically graded breast cancer, we have in this pilot project searched for additional protein biomarkers in a new space of the proteome. To this end, we have for the first time performed protein expression profiling of breast cancer tumor tissue, using recombinant antibody microarrays, targeting mainly immunoregulatory proteins. Thus, we have explored the immune system as a disease-specific sensor (clinical immunoproteomics). Uniquely, the results showed that several biologically relevant proteins reflecting histological grade could be delineated. In more detail, the tentative biomarker panels could be used to i) build a candidate model classifying grade 1 vs. grade 3 tumors, ii) demonstrate the molecular heterogeneity among grade 2 tumors, and iii) potentially re-classify several of the grade 2 tumors to more like grade 1 or grade 3 tumors. This could, in the long-term run, lead to improved prognosis, by which the patients could benefit from improved tailored care.

show abstract

Section: Methodsmentioning

confidence: 99%

Tumor tissue protein signatures reflect histological grade of breast cancer

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…The probe source (format) has been found to display superior on-chip performances compared with conventional monoclonal and polyclonal antibodies [25,26]. The specific 13 antibodies used in this study were selected based on the criteria that they were tested to work well in microarray applications and directed against target antigens located in the cell surface membrane [19,27,28]. More specifically, we use recombinant scFv antibodies that have been designed for microarray applications by molecular design, thus displaying high-on chip performance.…”

Section: Methodsmentioning

confidence: 99%

Interfacing Antibody-Based Microarrays and Digital Holography Enables Label-Free Detection for Loss of Cell Volume

et al. 2015

Self Cite

View full text Add to dashboard Cite

Background:We introduce the combination of digital holographic microscopy (DHM) and antibody microarrays as a powerful tool to measure morphological changes in specifically antibody-captured cells. The aim of the study was to develop DHM for analysis of cell death of etoposide-treated suspension cells.Results/methodology:We demonstrate that the cell number, mean area, thickness and volume were noninvasively measured by using DHM. The cell number was stable over time, but the two cell lines showed changes of cell area and cell irregularity after treatment. The cell volume in etoposide-treated cells was decreased, whereas untreated cells showed stable volume.Conclusion:Our results provide proof of concept for using DHM combined with antibody-based microarray technology for detecting morphological changes in captured cells.

show abstract

“…Examples include identification of disease signatures based on detection of multiple cytokines and inflammatory mediators [2][3][4]10,12,[20][21][22][23][24]. The results are commonly validated by independent assays such as ELISA and Western mediators [2][3][4]10,12,[20][21][22][23][24][25][26][27] (Table 1). In several cases, samples have been spiked with recombinant proteins to test specificity of the binders directly in the application (Table 1).…”

Section: Can Monospecific Binders Be Made?mentioning

confidence: 99%