Sergey N Krylov

• PhD
• Professor (Full) at York University

Maintaining stringent conditions in SELEX (Systematic Evolution of Ligands by EXponential enrichment) is crucial for obtaining high-affinity aptamers. However, excessive stringency greatly increases the risk of SELEX failure. Controlling stringency has remained a technical challenge, largely dependent on intuition, due to the absence of a clear, qu...

Activatable fluorescent probes with turn-on emission in response to a biological target can reduce the background signal and improve the detection limit needed for biosensor and bioimaging resolution. Turn-on probes with dual emission (ratiometric probes) have further advantages, as they have self-calibration ability for more reliable detection. He...

The equilibrium constant of affinity complexes (Kd) is among the most frequently determined physicochemical parameters, with thousands of papers reporting Kd values monthly. Kd is classically computed via nonlinear regression of a binding isotherm and, while it can be precise, it is often inaccurate due to error amplification. Currently, no method...

Heat shock protein 90 (HSP90) is a critical target for anticancer and anti-fungal-infection therapies due to its central role as a molecular chaperone involved in protein folding and activation. In this study, we developed in vitro Förster Resonance Energy Transfer (FRET) assays to characterize the binding of C. albicans HSP90 to its co-chaperone S...

The equilibrium dissociation constant (Kd) is a major characteristic of affinity complexes and one of the most frequently determined physicochemical parameters. Despite its significance, the values of Kd obtained for the same complex under similar conditions often exhibit considerable discrepancies and sometimes vary by orders of magnitude. These i...

Maintaining stringent conditions in SELEX (Systematic Evolution of Ligands by EXponential enrichment) is crucial for obtaining high-affinity aptamers; however, excessive stringency greatly increases the risk of SELEX failure. The control of stringency remains a technical challenge reliant solely on intuition, largely due to the absence of a measure...

High stringency (e.g., low target concentration) is key for obtaining high-affinity aptamers in SELEX; however, excessive stringency greatly increases the probability of SELEX failure. The control of stringency in SELEX remains a technical art based solely on intuition. A major reason for this is the lack of a measure of stringency. Here we introdu...

Non-steady-state behaviors are not expected in electric circuits that lack significant capacitance, inductivity, and/or active feedback. Here, we report that electrophoresis on paper (used to conduct/enhance paper-based assays) can create a non-steady-state electric circuit. We studied electrophoresis on 4-mm wide nitrocellulose-membrane strips uti...

The equilibrium dissociation constant (Kd) characterizes stability of binding complexes. The classic way of Kd determination involves finding the dependence of a fraction of unbound ligand on the total concentration of target (T0) when the total concentration of ligand (L0) remains constant. It is known that Kd determination for highly stable compl...

Intrinsically disordered proteins (IDPs), which can interact with many partner proteins, are central to many physiological functions and to various pathologies that include neurodegeneration. Here, we introduce the Sherpa hypothesis, according to which a subset of stable IDPs that we term Phenotype-Preserving Disordered Proteins (PPDP) play a centr...

Cytometry of Reaction Rate Constant (CRRC) is a method for studying cell-population heterogeneity using time-lapse fluorescence microscopy, which allows one to follow reaction kinetics in individual cells. The current and only CRRC workflow utilizes a single fluorescence image to manually identify cell contours which are then used to determine fluo...

The equilibrium dissociation constant (Kd) characterizes stability of non-covalent molecular complexes. Determining Kd for highly stable complexes may be extremely inaccurate if the ratio between the concentration of the limiting component (L0) and the a priori unknown value of Kd exceeds an unknown threshold value (aka threshold ratio). The only k...

The equilibrium dissociation constant (Kd) characterizes stability of non-covalent molecular complexes. Determining Kd for highly stable complexes may be extremely inaccurate if the ratio between the concentration of the limiting component (L0) and the a priori unknown value of Kd exceeds an unknown threshold value (aka threshold ratio). The only k...

Serological assays detect the presence of specific antibodies in blood. There are urgent and important applications for serological point-of-care (POC) assays. However, available detection methods are either insufficiently sensitive or too complex for POC settings. Here, we demonstrate that lateral flow immunoassay (LFIA), which is arguably the sim...

Intrinsically disordered proteins (IDPs) are central to many physiological functions and to various pathologies that include neurodegeneration. They can interact with numerous partner proteins. Here, we introduce the Sherpa hypothesis, according to which a subset of stable IDPs, which we term Phenotype-Preserving Disordered Proteins (PPDP), play a...

The determination of accurate equilibrium dissociation constants, Kd, of protein-small molecule complexes is important but challenging as all established methods have inherent sources of inaccuracy. Accurate Constant via Transient Incomplete Separation (ACTIS) is a new method for Kd determination using transient incomplete separation of the complex...

The determination of accurate equilibrium dissociation constants Kd of protein–small molecule complexes is important but challenging as all established methods have inherent sources of inaccuracy. ACTIS is a new method for Kd determination using transient incomplete separation of the complex from the unbound small molecule in a pressure-driven flow...

Lateral flow immunoassay (LFIA) is a rapid, simple, and inexpensive point‐of‐need method. A major limitation of LFIA is a high limit of detection (LOD), which impacts its diagnostic sensitivity. To overcome this limitation, we introduce a signal‐enhancement procedure that is performed after completing LFIA and involves controllably moving biotin‐ a...

Lateral flow immunoassay (LFIA) is a rapid, simple, and inexpensive point‐of‐need method. A major limitation of LFIA is a high limit of detection (LOD), which impacts its diagnostic sensitivity. To overcome this limitation, we introduce a signal‐enhancement procedure that is performed after completing LFIA and involves controllably moving biotin‐ a...

Large molecules can be generically separated from small ones, though partially and temporarily, in a pressure-driven flow inside a capillary. This transient incomplete separation has been only applied to species with diffusion coefficients different by at least an order of magnitude. Here, we demonstrate, for the first time, the analytical utility...

Selection of oligonucleotide aptamers involves consecutive rounds of affinity isolation of target-binding oligonucleotides from a random-sequence oligonucleotide library. Every next round produces an aptamer-enriched library with progressively higher fitness for tight binding to the target. The progress of enrichment can only be accurately assessed...

A low-cost and label-free microfluidic resistive biosensor was developed for bacteria detection. The proposed design employed two gold-coated copper microbridge wires suspended orthogonally in a PDMS microchannel. Anti-Escherichia coli (E. coli) antibodies were immobilized on the wires for capturing them from the water sample. Electrical resistance...

A low-cost and handheld thermo-photonic biosensor was developed for bacteria detection. Anti-Escherichia (E.) Coli antibodies were immobilized on gold-coated wires for capturing E. coli K12 from the water sample. Then, the captured bacteria were sandwiched with gold nanoparticles conjugated with antibodies. Following laser excitation, the developed...

Lateral flow immunoassay (LFIA) is a rapid, simple, and inexpensive method for point-of-need analysis. A major limitation of LFIA is a high limit of detection (LOD), which impacts its diagnostic sensitivity. To overcome this limitation, we introduce a signal-enhancement procedure that is performed after completing LFIA and involves controllably mov...

Molecular stream separation (MSS) is a promising complement for continuous-flow synthesis. MSS is driven by forces exerted on molecules by a field applied at an angle to the stream-carrying flow. MSS has only been performed with a 90° field-to-flow angle because of a rectangular geometry of canonic MSS; the second-order rotational symmetry of a rec...

Cytometry of Reaction Rate Constant (CRRC) is a method for studying heterogeneity of cell populations with regards to activity of cellular reactions. It is based on time-lapse fluorescence microscopy which facilitates following reaction kinetics in individual cells. The current CRRC workflow utilizes a single fluorescence image to manually identify...

Molecular stream separation (MSS) is a promising complement for continuous-flow synthesis. MSS is driven by forces exerted on molecules by a field applied at an angle to the stream-carrying flow. MSS has only been performed with a 90 field-to-flow angle because of a rectangular geometry of canonic MSS; the second-order rotational symmetry of a rec...

Molecular stream separation (MSS) is a promising complement for continuous-flow synthesis. MSS is driven by forces exerted on molecules by a field applied at an angle to the stream-carrying flow. MSS has only been performed with a 90 field-to-flow angle because of a rectangular geometry of canonic MSS; the second-order rotational symmetry of a rec...

Partitioning of protein–DNA complexes from protein-unbound DNA is a key step in selection of DNA aptamers. Conceptually, the partitioning step is characterized by two parameters: transmittance for protein-bound DNA (binders) and transmittance for unbound DNA (nonbinders). Here we present the first study to reveal how these transmittances depend on...

Partitioning of protein–DNA complexes from protein-unbound DNA is a key step in selection of DNA aptamers. Conceptually, the partitioning step is characterized by two parameters: transmittance for protein-bound DNA (binders) and transmittance for unbound DNA (nonbinders). Here we present the first study to reveal how these transmittances depend on...

Accurate Constant via Transient Incomplete Separation (ACTIS) is a new method for finding the equilibrium dissociation constant Kd of a protein-small molecule complex based on transient incomplete separation of the complex from the unbound small molecule in a capillary. This separation is caused by differential transverse diffusion of the complex a...

In molecular-stream separation (MSS), a stream of a multicomponent mixture is separated into multiple streams of individual components. Quantitative evaluation of MSS data has been a bottleneck in MSS for decades as there was no conventional way to present the data in a reproducible and uniform fashion. The roots of the problem were in the multidim...

A primary reason for chemotherapy failure is chemoresistance, which is driven by various mechanisms. Multi-drug resistance (MDR) is one such mechanism that is responsible for drug extrusion from the intracellular space. MDR can be intrinsic and thus, may pre-exist the first application of chemotherapy. However, MDR may also be acquired during tumor...

p>ACTIS is a new method for finding the equilibrium dissociation constant K _d of a protein–small molecule complex based on transient incomplete separation of the complex from the unbound small molecule in a capillary. This separation is caused by differential transverse diffusion of the complex and the small molecule in a pressure-driven...

Screening molecular libraries for ligands capable of binding proteins is widely used for hit identification in the early drug discovery process. Oligonucleotide libraries provide a very high diversity of compounds, while the combination of the polymerase chain reaction and DNA sequencing allow the identification of ligands in low copy numbers selec...

p> In molecular-stream separation (MSS), a stream of a multi-component mixture is separated into multiple streams of individual components. Quantitative evaluation of MSS data has been a bottleneck in MSS for decades as there was no conventional way to present the data in a reproducible and uniform fashion. The roots of the problem were in the mult...

Thousands of putative microRNA (miRNA)-based cancer biomarkers have been reported, but none has been validated for approval by the Food and Drug Administration. One of the reasons for this alarming discrepancy is the lack of a method that is sufficiently robust for carrying out validation studies, which may require analysis of samples from hundreds...

Enzymes of the cytochrome P450 (CYP) family catalyze the metabolism of chemotherapeutic agents and are among the key players in primary and acquired chemoresistance of cancer. The activity of CYP is heterogeneous in tumor tissues, and the quantitative characteristics of this heterogeneity can be used to predict chemoresistance. Cytometry of reactio...

Cytometry of reaction rate constant (CRRC) is an accurate and robust approach to characterize cell-population heterogeneity using rate constants of cellular processes for which kinetic mechanisms are known. We work on a CRRC-based method to develop predictors of tumor chemoresistance driven by two processes: drug extrusion by multi-drug-resistance...

Here we present the result of our work on achieving sub-pM limit of quantitation in direct quantitative analysis of multiple miRNAs by capillary electophoresis with laser-induced fluorescence detection. The PDF file contains the description of the results with figures, references, and supporting information. The raw data.ZIP file is the archive con...

Chemoresistance, i.e., tumor insensitivity to chemotherapy, shortens life expectancy of cancer patients. Despite the availability of new treatment options, initial systemic regimens for solid tumors are dominated by a set of standard chemotherapy drugs, and alternative therapies are used only when a patient has demonstrated chemoresistance clinical...

Analytical methods may not have reference standards required for testing their accuracy. We postulate that accuracy of an analytical method can be assessed in the absence of reference standards in silico if the method is built upon deterministic processes. A deterministic process can be precisely computer-simulated thus allowing virtual experiments...

Does cell clustering influence intrinsic and acquired multi-drug resistance (MDR) differently? To address this question, we studied cultured monolayers (representing individual cells) and cultured spheroids (representing clusters) formed by drug-naïve (intrinsic MDR) and drug-exposed (acquired MDR) lines of ovarian cancer A2780 cells by cytometry o...

Does cell clustering in ovarian cancer influence intrinsic and acquired multi-drug resistance (MDR) differently? To address this question, we studied cultured monolayers (representing individual cells) and cultured spheroids (representing clusters) formed by drug-naive (intrinsic MDR) and drug-exposed (acquired MDR) lines of ovarian cancer A2780 ce...

Cytometry of Reaction Rate Constant (CRRC) uses time-lapse fluorescence microscopy to measure a rate constant of a catalytic reaction in individual cells and, thus, facilitate accurate size determination for cell subpopulations with distinct efficiencies of this reaction. Reliable CRRC requires uniform exposure of cells to the reaction substrate fo...

p>Analytical methods may not have reference standards required for testing their accuracy. We postulate that accuracy of an analytical method can be assessed in the absence of reference standards in silico if the method is built upon deterministic processes. A deterministic process can be precisely computer-simulated thus allowing virtual experimen...

File main pdf file (CRRC of dispersed-settled spheroidal cells.pdf) describes experimental results and their interpretation for a study of tissue relevance of analyses of dispersed-settled tissues cells by Cytometry of Reaction Rate Constant (CRRC). CRRC uses time-lapse fluorescence microscopy to measure a rate constant of a catalytic reaction in i...

Ideal‐filter capillary electrophoresis (IFCE) is a method for the selection of affinity binders for protein targets from oligonucleotide libraries, e.g., random‐sequence oligonucleotide libraries and DNA‐encoded libraries, in a single step of partitioning. In IFCE, protein–oligonucleotide complexes and unbound oligonucleotides move in the opposite...

Continuous-flow electrophoresis (CFE) separates a stream of a multi-component mixture into multiple streams of individual components inside a thin rectangular chamber. CFE will be able to benefit flow chemistry when it is both compatible with non-aqueous solvents utilized in organic synthesis and capable of generically detecting streams of small or...

Continuous-flow electrophoresis (CFE) separates a stream of a multi-component mixture into multiple streams of individual components inside a thin rectangular chamber. CFE will be able to benefit flow chemistry when it is capable of generically detecting streams of small organic molecules. Here we propose a general approach for molecular stream vis...

DNA aptamers are single-strand DNA (ssDNA) capable of selectively and tightly binding a target molecule. Capillary electrophoresis-based selection of aptamers for protein targets requires the knowledge of electrophoretic mobilities of protein–aptamer complexes while measuring these mobilities requires having the aptamers. Here we report on breaking...

Continuous-flow organic synthesis naturally requires continuous-flow separation of reaction components. The most common continuous-flow separation approach is liquid-liquid extraction based on differential distribution of molecules between organic and aqueous phases. This approach has limited selectivity; it can hardly separate different hydrophobi...

Ideal-filter capillary electrophoresis (IFCE) allows selection of protein binders from oligonucleotide libraries in a single step of partitioning in which protein–bound and unbound oligonucleotides move in the opposite directions. In IFCE, the unbound oligonucleotide does not reach the detector, imposing a problem for finding the equilibrium consta...

Selection of affinity ligands for protein targets from oligonucleotide libraries currently involves multiple rounds of alternating steps of partitioning of protein‐bound oligonucleotides (binders) from protein‐unbound oligonucleotides (nonbinders). We have recently introduced Ideal‐Filter Capillary Electrophoresis (IFCE) for binder selection in a s...

The pdf file describes the results of this study, which aims at developing a model to accurately predict electrophoretic mobilities of protein–aptamer complexes. The excel file contains source data for electropherograms (signal vs time) which were obtained in this study and used to determine the migration times and electrophoretic mobilities of pro...

Current practical methods for finding the equilibrium dissociation constant, Kd, of protein–small molecule complexes have inherent sources of inaccuracy. Introduced here is “accurate constant via transient incomplete separation” (ACTIS), which appears to be free of inherent sources of inaccuracy. Conceptually, a short plug of the pre‐equilibrated p...

div>Continuous-flow organic synthesis naturally requires continuous-flow separation of reaction components. The most common continuous-flow separation approach is liquid-liquid extraction based on differential distribution of molecules between organic and aqueous phases. This approach has limited selectivity; it can hardly separate different hydrop...

Robust and accurate analysis of cell-population heterogeneity is challenging but required in many areas of biology and medicine. In particular, it is pivotal to the development of reliable cancer biomarkers. Here, we prove that cytometry of reaction rate constant (CRRC) can facilitate such analysis when the kinetic mechanism of a reaction associate...

Current practical methods for finding the equilibrium dissociation constant, Kd, of protein–small molecule complexes are mainly biosensoric and calorimetric. They have inherent sources of inaccuracy; immobilization of molecules on sensors and heat of side reactions often lead to dramatic errors in Kd. We introduce “accurate constant via transient i...

Selection of aptamers from oligonucleotide libraries currently requires multiple rounds of alternating steps of partitioning of binders from nonbinders and enzymatic amplification of all collected oligonucleotides. Herein, we report a highly practical solution for reliable one‐step selection of aptamers. We introduce partitioning by ideal‐filter ca...

div>Current methods for finding the equilibrium dissociation constant, K _d, of protein-small molecule complexes have inherent sources of inaccuracy. We introduce “Accurate K _d via Transient Incomplete Separation” (AKTIS), an approach that is free of known sources of inaccuracy. Conceptually, in AKTIS, a short plug of the pre-...

Selection of aptamers from oligonucleotide libraries currently requires multiple rounds of alternating steps of partitioning of binders from nonbinders and enzymatic amplification of all collected oligonucleotides. Here we report a highly‐practical solution for reliable one‐step selection of aptamers. We introduce partitioning by Ideal‐Filter Capil...

Direct quantitative analysis of multiple miRNAs (DQAMmiR) is a hybridization-based assay in which the excess of the DNA hybridization probes is separated from the miRNA-probe hybrids and the hybrids are separated from each other in gel-free capillary electrophoresis (CE) using two types of mobility shifters: single-strand DNA binding protein (SSB)...

Circular RNAs are a large group of noncoding RNAs that are widely expressed in mammalian cells. Genome-wide analyses have revealed abundant and evolutionarily conserved circular RNAs across species, which suggest specific physiological roles of these species. Using a microarray approach, we detected increased expression of a circular RNA circ-Dnmt1...

Determining kinetic rate constants is a highly relevant problem in biochemistry, so various methods have been designed to extract them from experimental data. Such methods have two main components: the experimental apparatus and the subsequent analysis, the latter dependent on the mathematical approach taken, which influences the effectiveness of c...

We propose Cytometry of Reaction Rate Constant (CRRC) for accurate analysis of cell-population heterogeneity with respect to a specific molecular reaction. Conceptually, in CRRC, the cells are loaded with a reaction substrate, and its conversion into a product is followed by time-lapse fluorescence microscopy at the single-cell level. A reaction ra...

Nonequilibrium capillary electrophoresis of equilibrium mixtures (NECEEM) is an affinity method for separating binder-target complexes from nonbinders by gel-free CE. NECEEM is a promising high-efficiency method for partitioning protein binders from nonbinders in DNA-encoded libraries (DEL), such binders are used as "hits" in drug development. It i...

Molecular-stream separation (MSS), e.g. by free flow electrophoresis or continuous annular chromatography, has great potential for applications that require continuous downstream separation such continuous flow synthesis. Despite its potential, MSS still needs to be greatly advanced, which requires currently lacking tools for quantitative character...

miRNAs play important roles in gene regulation, and their dysregulation is associated with many diseases, including epithelial ovarian cancer (EOC). In this study, we determined the expression and function of miR-590-3p in EOC. miR-590-3p levels were higher in high-grade carcinoma when compared with low-grade or tumors with low malignant potential....

Molecular-stream separation (MSS), e.g. by free flow electrophoresis or continuous annular chromatography, has great potential for applications that require continuous downstream separation such continuous flow synthesis. Despite its potential, MSS still needs to be greatly advanced, which requires currently lacking tools for quantitative character...

Molecular-stream separation (MSS), e.g. by free flow electrophoresis or continuous annular chromatography, has great potential for applications that require continuous downstream separation such continuous flow synthesis. Despite its potential, MSS still needs to be greatly advanced, which requires currently lacking tools for quantitative character...

DNA aptamers are attractive capture probes for affinity chromatography since, in contrast to antibodies, they can be chemically synthesized and, in contrast to tag-specific capture probes (such as Nickel-NTA or Glutathione), they can be used for purification of proteins free of genetic modifications (such as His or GST tags). Despite these attracti...

Here we introduce Protein-Lipidation Quantitation (PLQ) – the first method for quantitative analysis of both a substrate and a product of protein lipidation in a biologically relevant context. Such analysis is required to study roles of protein lipidation in cellular regulation. In PLQ, the substrate is fused with a fluorescent protein to facilitat...

The limit-of-detection (LOD) in analytical instruments with fluorescence detection can be improved by reducing noise of optical background. Efficiently reducing optical background noise in systems with spectrally non-uniform background requires complex optimization of an emission filter – the main element of spectral filtration. Here we introduce a...

A discretization scheme is introduced for a set of convection–diffusion–reaction equations with a non-linear reaction term describing reversible binding and a constant convection velocity for each reactant. This constancy of convection velocities allows a transformation to new spatial variables, which ensures the global stability of discretization....

Analysis of proteinogenic vaccine antigens in a quality control environment requires an accurate, precise, and reliable method for protein separation and quantitation. While having multiple advantages over the classical SDS-PAGE, capillary gel electrophoresis (CGE) has not yet become a standard tool in vaccine antigen analysis. Here we report on de...

The construction of switchable, radiation-controlled, aptameric enzymes - "swenzymes" - is, in principle, feasible. We propose a strategy to make such catalysts from 2 (or more) aptamers each selected to bind specifically to one of the substrates in, for example, a 2-substrate reaction. Construction of a combinatorial library of candidate swenzymes...

Aims: Clear cell renal cell carcinoma (ccRCC) is the most common adult kidney cancer. It is an aggressive tumour with unpredictable outcome. The currently used clinical parameters are not always accurate for predicting disease behaviour. miR-10b is dysregulated in different malignancies including RCC. Methods: We assessed the clinical utility of...

Accurate quantitation of microRNA (miRNA) in tissue samples is required for validation and clinical use of miRNA-based disease biomarkers. Since sample processing, such as RNA extraction, introduces undesirable biases, it is advantageous to measure miRNA in a crude cell lysate. Here we report on accurate miRNA quantitation in crude cell lysate by a...

We present an image processing and analysis system to facilitate detailed performance analysis of free flow electrophoresis (FFE) chips. It consists of a cost-effective self-built imaging setup and a comprehensive customizable software suite. Both components were designed modularly to be accessible, adaptable, versatile, and automatable. The system...

A discretization scheme is introduced for a set of convection-diffusion equations with a non-linear reaction term, where the convection velocity is constant for each reactant. This constancy allows a transformation to new spatial variables, which ensures the global stability of discretization. Convection-diffusion equations are notorious for their...

Here we report on unexpected electrophoretic behavior of complexes between a rod-like virus particles (virions) and bivalent antibodies. The multiple complexes formed by the virions and antibodies migrated with electrophoretic mobilities of much greater absolute values than those of the unbound virions or antibody while typically complexes have mob...

Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures (NECEEM) is an efficient method for studying intermolecular interactions. Optimization of NECEEM experiments is not a trivial task, due to the complex interrelation between numerous experimental parameters and their combined effects on the accuracy and precision of measurements. Here...

In this technical note, we demonstrate the hyphenation of production-scale free-flow electrophoresis (FFE) and sheathless electrospray ionization mass spectrometry (ESI-MS). In contrast to previous hyphenation approaches, we used a highly conductive background electrolyte (BGE) required for production-scale FFE. We found that this kind of BGE as we...

Kinetic Capillary Electrophoresis (KCE) methods are useful in the study of kinetics and equilibrium properties of interactions between DNA and its binding partners (ligands). KCE experiments are typically performed in a narrow set of "conventional" low-conductivity run buffers while DNA-ligand interactions in biological systems occur in physiologic...

In functional cytometric studies, cultured cells are exposed to effectors (e.g. drugs), and the heterogeneity of cell responses are studied using cytometry techniques (e.g. image cytometry). Such studies are difficult to perform on 3D cell cultures. A solution is to disperse 3D clusters and transfer the cells to the 2D state before applying effecto...

Capillary electrophoresis with Laser-Induced Fluorescence (CE-LIF) detection is being applied to new analytical problems which challenge both the power of CE separation and the sensitivity of LIF detection. On-capillary LIF detection is much more practical than post-capillary detection in a sheath-flow cell. Therefore, commercial CE instruments uti...

Selection of target-binding ligands from DNA-encoded libraries of small molecules (DELSMs) is a rapidly developing approach in drug-lead discovery. Methods of kinetic capillary electrophoresis (KCE) may facilitate highly efficient homogeneous selection of ligands from DELSMs. However, KCE methods require accurate prediction of electrophoretic mobil...

MicroRNAs (miRNAs) and piwi-interacting RNAs (piRNAs) are two classes of small noncoding RNAs, both of which play roles in regulating tissue development. It is unknown whether these distinct classes of noncoding RNAs can regulate one another. Here we show that ectopic expression of miR-17 inhibited mouse fertility and early embryonic development. S...

Kinetic size exclusion chromatography with mass-spectrometry detection (KSEC-MS) is a solution-based label-free approach for studying kinetics of reversible binding of a small molecule to a protein. Extraction of kinetic data from KSEC-MS chromatograms is greatly complicated by the lack of separation between the protein and protein-small molecule c...