Raising antibodies against circulating foetal cells from maternal peripheral blood

Sørensen, Morten Dræby; Melchjorsen, Connie Jenning; Mandrup, Ole Aalund; Kristensen, Peter

doi:10.1002/pd.4060

Cited by 4 publications

(6 citation statements)

References 51 publications

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“…The present section describes antibody selection on whole FFPE sections [57,58], and also three on-slide micro-selection strategies to target specific tissue structures or cell clusters (Figure 1). Two of them rely on the excision of the positive tissue by laser-assisted microdissection [59][60][61][62][63] or with a micropipette [64]; the third one blocks bacterial replication of irrelevant phages through UV-induced DNA damage [65][66][67][68][69][70][71]. All these strategies can be used on FFPE and frozen tissue sections.…”

Section: On-slide Antibody Selectionmentioning

confidence: 99%

“…One of the characteristics of the shadow stick method is to "provide few and highly relevant output clones" [65]. The number of selected clones is low enough to allow their direct screening: eight clones per selection after optimization in 2010 [66]; 12 clones in three selections in 2013 [68]; 40 clones in two selections on FFPE sections in 2015 [65]; 315 clones in 13 selections on frozen sections in 2015 and 2016 [69,70]; and 93 clones in 8 selections on frozen sections in 2017 [71]. This low clone number greatly facilitates the subsequent screening steps.…”

Section: Shadow Stick-based Antibody Selectionmentioning

confidence: 99%

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Antibody Identification for Antigen Detection in Formalin-Fixed Paraffin-Embedded Tissue Using Phage Display and Naïve Libraries

Mairaville¹,

Martineau²

2021

Antibodies

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Immunohistochemistry is a widely used technique for research and diagnostic purposes that relies on the recognition by antibodies of antigens expressed in tissues. However, tissue processing and particularly formalin fixation affect the conformation of these antigens through the formation of methylene bridges. Although antigen retrieval techniques can partially restore antigen immunoreactivity, it is difficult to identify antibodies that can recognize their target especially in formalin-fixed paraffin-embedded tissues. Most of the antibodies currently used in immunohistochemistry have been obtained by animal immunization; however, in vitro display techniques represent alternative strategies that have not been fully explored yet. This review provides an overview of phage display-based antibody selections using naïve antibody libraries on various supports (fixed cells, dissociated tissues, tissue fragments, and tissue sections) that have led to the identification of antibodies suitable for immunohistochemistry.

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Section: On-slide Antibody Selectionmentioning

confidence: 99%

Section: Shadow Stick-based Antibody Selectionmentioning

confidence: 99%

Antibody Identification for Antigen Detection in Formalin-Fixed Paraffin-Embedded Tissue Using Phage Display and Naïve Libraries

Mairaville¹,

Martineau²

2021

Antibodies

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“…The method has been dubbed ‘shadow-stick selection’ due to the use a minute disk attached to a glass pipette (resembling a hockey-stick), which is used to selectively protect the target cell, and the phage bound to it, from UV irradiation. By reducing the requirement to a single identified cell it was possible to select antibodies against an individual cultured K562 cell spiked into peripheral blood mononuclear cells (PBMC) [32] and against cells identified as fetal microchimeric cells in maternal blood [23]. This method is especially useful if the target cell is present in a very low frequency.…”

Section: Ex Vivo Phage Selectionmentioning

confidence: 99%

Selection strategies for anticancer antibody discovery: searching off the beaten path

Sánchez-Martín¹,

Sørensen²,

Lykkemark³

et al. 2015

Trends in Biotechnology

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Antibody based drugs represent one of the most successful and promising therapeutic approaches in oncology. Large combinatorial phage antibody libraries are available for identification of therapeutic antibodies, and a variety of technologies exist for their further conversion into multivalent and multispecific formats optimized for the desired pharmacokinetics and the pathological context. However, there is no technology for antigen profiling of intact tumors to identify tumor markers targetable with antibodies. Such constraints have led to a relative paucity of tumor-associated antigens for antibody targeting in oncology. Here we review novel approaches aimed at the identification of antibody-targetable, accessible antigens in intact tumors. We hope that such advanced selection approaches will be useful in the development of next-generation antibody therapeutics for cancer.

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“…Phage display-mediated selections of human domain antibodies were performed against ductal regions containing K19 + /K14 + cells in normal breast tissue cryostat sections using the shadow stick selection technique [15, 16] adopted from [17–19]. Here, a recombinant antibody library, displayed in genetic fusion with the filamentous bacteriophage protein 3 [20], is allowed to bind to antigens presented on the tissue sections.…”

Section: Introductionmentioning

confidence: 99%

Upregulation of Mrps18a in breast cancer identified by selecting phage antibody libraries on breast tissue sections

et al. 2017

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BackgroundOne of the hallmarks of cancer is an altered energy metabolism, and here, mitochondria play a central role. Previous studies have indicated that some mitochondrial ribosomal proteins change their expression patterns upon transformation.MethodIn this study, we have used the selection of recombinant antibody libraries displayed on the surface of filamentous bacteriophage as a proteomics discovery tool for the identification of breast cancer biomarkers. A small subpopulation of breast cells expressing both cytokeratin 19 and cytokeratin 14 was targeted using a novel selection procedure.ResultsWe identified the mitochondrial ribosomal protein s18a (Mrps18a) as a protein which is upregulated in breast cancer. However, Mrps18a was not homogeneously upregulated in all cancer cells, suggesting the existence of sub-populations within the tumor. The upregulation was not confined to cytokeratin 19 and cytokeratin 14 double positive cells.ConclusionThis study illustrates how phage display can be applied towards the discovery of proteins which exhibit changes in their expression patterns. We identified the mitochondrial protein Mrps18a as being upregulated in human breast cancer cells compared to normal breast cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2987-5) contains supplementary material, which is available to authorized users.

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Raising antibodies against circulating foetal cells from maternal peripheral blood

Abstract: These findings indicate that the identified foetal cells could have been progenitor cells of haematopoietic origin. The antibody SF1.3 could be a potential tool toward non-invasive prenatal diagnostics.

Cited by 4 publications

References 51 publications

Antibody Identification for Antigen Detection in Formalin-Fixed Paraffin-Embedded Tissue Using Phage Display and Naïve Libraries

Antibody Identification for Antigen Detection in Formalin-Fixed Paraffin-Embedded Tissue Using Phage Display and Naïve Libraries

Selection strategies for anticancer antibody discovery: searching off the beaten path

Upregulation of Mrps18a in breast cancer identified by selecting phage antibody libraries on breast tissue sections

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