Oda Stoevesandt's research while affiliated with Babraham Institute and other places

... Cross-reactivity of affinity reagents limits the number of proteins that can be evaluated in a multiplexed immunoaffinity-based assay to the low hundreds (2). Single-stranded DNA aptamerbased assays have been developed as an alternative to immunoaffinity-based methods (3) and have been increasingly applied in clinical and epidemiological studies (4)(5)(6)(7)(8). For example, a panel of 9 proteins identified from thousands has been shown to predict cardiovascular disease better than traditional risk factors (4). ...

... TAP requires that the interactions are sufficiently stable to survive the isolation process and low-level expressed complexes may be missed. A third group of methods based on partner proximity use labelled detection reagents such as antibodies to localise proteins in close apposition to each other, by the interaction of the labels, for example, fluorescence cross-correlation spectroscopy (FCCS) [9], fluorescence resonance energy transfer (FRET) [10] or oligonucleotide ligation followed by PCR or rolling circle amplification (proximity ligation) [11,12]. These methods, which may be intracellular or solution based, depend on the availability of specific detection reagents to identify pairwise interactions and usually assume some knowledge of the identity of the interaction partners. ...

... Similarly, coupling to the HPMA polymer afforded a proteolytic stabilization of a peptide derived from the interaction motif for an SH3 domain. Following the import into cells, the synthetic scaffold improved the formation of microclusters of signaling proteins [30]. ...

... To develop such array designs, several fabrication techniques have been explored as these are key to their overall performance and reliability. 6 These range from conventional contact 16−18 and noncontact-based printing 19−21 to lithography 22−24 and microfluidics, 25−27 which are as diverse as cell-free protein expression-based printing. 28 Printing reproducibility has been one of the most crucial issues affecting the traditional protein chips. ...

... The international community has also come up with a proposal called Minimum Information About a Protein Affinity Reagent (MIAPAR), which aims to establish a stronger connection between antibody producers and users. MIAPARcompliant data includes information such as the production/purification process, experimental evidence, updated protocols, and other relevant details (7,9). ...

... In terms of number of publications, NAPPA from the LaBear group can be accounted as leading method [9,10,[19][20][21][62][63][64][65][66][67][68][69][70][71][72][73][74][75][76][77][78][79][80], followed by DAPA [28,29,[81][82][83][84][85][86][87], and PING [32,[88][89][90][91][92][93]. DAPA, pMAC, and μIP offer the opportunity to make multiple copies from a template DNA microarray, which is not possible by NAPPA. ...

... The international community has also come up with a proposal called Minimum Information About a Protein Affinity Reagent (MIAPAR), which aims to establish a stronger connection between antibody producers and users. MIAPAR-compliant data include information such as the production/purification process, experimental evidence, updated protocols, and other relevant details (7,9). ...

... The proteins once expressed become rapidly immobilized on the capture slide, creating a protein array that can be duplicated several times. DAPA has been extended to print arrays of 116 different proteins (Stoevesandt, Vetter et al. 2011). It has also been optimized using different array support coatings, which increase its sensitivity towards the printed proteins (Schmidt, Cook et al. 2013). ...

... Using this approach, which begins with a library of single-chain variable fragments (scFv's) containing an engineered anion-binding pocket (or "nest") and then uses phage display to enrich for scFv's that recognize a specific immobilized phosphopeptide, the authors were able to identify >50 antibodies that exhibit high specificity for the phosphorylated forms of their respective targets. This generalizable approach has the potential to dramatically expand the toolkit of reliable phospho-specific antibodies, which will not only facilitate the development of analytical protein microarrays but also promises to increase the utility of other approaches based on immunological detection of phosphorylated residues (Stoevesandt & Taussig, 2013). ...

... The phage display technology was further developed and improved by the following research teams: G. Winter [6][7][8][9][10][11]. All these researchers pursued the creation of phage-displayed combinatorial antibodies libraries, which were further improved by several other laboratories in the following years [12][13][14][15][16][17][18][19][20]. As recognition of the phage display contribution to scientific advances in chemistry and pharmaceutics, George P. Smith and Sir Gregory P. Winter received the 2018 Nobel Prize in Chemistry "for phage visualization of peptides and antibodies" [21]. ...