Emilio Margolles-clark - Academia.edu (original) (raw)

Papers by Emilio Margolles-clark

Cloning of genes encoding alpha-L-arabinofuranosidase and beta-xylosidase from Trichoderma reesei... more Cloning of genes encoding alpha-L-arabinofuranosidase and beta-xylosidase from Trichoderma reesei by expression in Saccharomyces cerevisiae.

Transplantation Pathology

Human Immunology, 2018

Aim: Our lab has established the NXType TM kit on the Ion-Chef and Ion-S5 workflow for high resol... more Aim: Our lab has established the NXType TM kit on the Ion-Chef and Ion-S5 workflow for high resolution HLA typing. This work was aimed to evaluate the analytical performance of the method for accuracy, precision, sensitivity, specificity, reportable range, repeatability, and reproducibility. We also determined optimal run size and workflow within the lab to strike a balance between suitable number of reads per locus and number of samples to avoid low coverage and dropouts. Using NGS as primary typing method for hematopoietic stem cell (HSC) and the solid organ (SO) programs was assessed. Methods: Class I and HLA Class II were amplified with NXType TM or ALLType TM primer sets. Automated template preparation and chip loading were performed in the Ion Chef TM system and run on Ion S5 TM. Data was analyzed with TypeStream TM v1.1.0.11 and TypeStream TM Visual NGS Analysis Software. Results: We found that the optimal run size was a minimum of 24 samples, a 3-fold increase of previous capacity. To maximize the use of chip space the number of samples/run was increased with confirmatory solid organ typings. We determined that running 24 samples with SSOP or NGS required similar manpower and instrument time. Although lab work was less with SSOP, analysis time and effort was greater than with NGS. After two years of experience with NGS HLA typing (1500 samples), we considered using NGS as the primary typing method for the HSC and the solid organ programs. Our historical data showed only 2 cases where NGS gave non-concordant results with SSOP or SSP. However, these samples were flagged by the software and both had associations that were red-flagged during analysis. The repeat rate for SSOP was 2.8% while for NGS was 0.3%. All NGS repeats were technical failures and not ambiguity resolution. We evaluated the ALLtype kit, which includes DQA1 necessary for solid organ initial typing, and a liquid handler for automation of library preparation. In addition, we implemented the TypeStream Visual (TSV) analysis software that includes serological equivalents. Conclusions: We confirmed that NGS is a robust methodology for HLA typing. It provides results first time is run, reducing reflex testing. Combining automation with ALLType TM kit and TSV software, the analysis will be used as the primary method and SSOP as a reflex method for HSC and SO.

Applied and Environmental Microbiology, 1996

The chromosomal endochitinase gene (ThEn-42) of the mycoparasite fungus Trichoderma harzianum P1 ... more The chromosomal endochitinase gene (ThEn-42) of the mycoparasite fungus Trichoderma harzianum P1 was isolated and overexpressed in the filamentous fungus Trichoderma reesei under the promoter of the major cellulase gene cbhl1. The host strain RutC-30 did not produce any endogenous endochitinase activity. The prepro region of the T harzianum endochitinase was correctly processed in T. reesei. No differences in expression were observed when the prepro region was replaced with the CBHI signal sequence. Shake flask cultivation yielded 130 mg of active enzyme per liter, which in terms of activity represents about a 20-fold increase over the endochitinase activity produced by T. harzianum. The presence of multiple copies of the expression cassette in the transformant resulted in limitation in transcription and/or regulation factors needed for full activity of the cbh1 promoter, although this was not the major limiting factor for higher expression of endochitinase. The endochitinase was ve...

Applied and environmental microbiology, 1996

Production of extracellular endochitinase could be increased 5-fold in the mycoparasite fungus Tr... more Production of extracellular endochitinase could be increased 5-fold in the mycoparasite fungus Trichoderma harzianum by using the cellulase promoter cbh1 of Trichoderma reesei, whereas the total endochitinase activity increased 10-fold. The cbh1 promoter was not expressed on glucose and sucrose in T. harzianum and was induced by sophorose and on cellulase-inducing medium. The endogenous endochitinase gene was expressed at a low basal level on glucose and sucrose. No specific induction by crab shell chitin or sophorose was observed.

Applied and Environmental Microbiology, 1996

A cDNA expression library of Trichoderma reesei RutC-30 was constructed in the yeast Saccharomyce... more A cDNA expression library of Trichoderma reesei RutC-30 was constructed in the yeast Saccharomyces cerevisiae. Two genes, abf1 and bxl1, were isolated by screening the yeast library for extracellular alpha-L-arabinofuranosidase activity with the substrate p-nitrophenyl-alpha-L-arabinofuranoside. The genes abf1 and bxl1 encode 500 and 758 amino acids, respectively, including the signal sequences. The deduced amino acid sequence of ABFI displays high-level similarity to the alpha-L-arabinofuranosidase B of Aspergillus niger, and the two can form a new family of glycosyl hydrolases. The deduced amino acid sequence of BXLI shows similarities to the beta-glucosidases grouped in family 3. The yeast-produced enzymes were tested for enzymatic activities against different substrates. ABFI released L-arabinose from p-nitrophenyl-alpha-L-arabinofuranoside and arabinoxylans and showed some beta-xylosidase activity toward p-nitrophenyl-beta-D-xylopyranoside. BXLI did not release L-arabinose from...

Human Immunology, 2019

The common methods for HLA typing in cadaveric donors are low to intermediate resolution (i.e SSP... more The common methods for HLA typing in cadaveric donors are low to intermediate resolution (i.e SSP and/ or SSOP) due to time-constraints. Both methods may fail to distinguish among different alleles in the target region and/or alleles lying outside the amplified region. Our lab has established the Ion-Torrent NGS workflow for confirmatory high resolution HLA typing in cadaveric donors. Here we present a case where typing by SSP (LinkSeq TM) resulted in the allele combination A*24/A*31 with only well-documented (d) or rare (r) alleles associated with the A*24. Because of the ''r" alleles, typing was reflexed to SSOP. The results indicated the most probable allele combination would be A*24:02/A*31:01 Final serological typing was reported to UNet as A24/A31. Subsequently, confirmatory typing was performed by NGS. The results showed the typing as A*24:53/A*31:01. Although present in the string of both LinkSeq and SSOP, this result was not the expected typing. Additionally, the nomenclature varied between the LinkSeq and the SSOP results. LinkSeq considered the A*24:53 as a 'd' allele, or not common. SSOP considered the combination as 'frequent on both alleles' or G1. This case shows evidence that the most common or 'expected' allele combination is not always the correct one. It is important to keep in mind the purpose of the HLA typing. In this case, a potential solid organ donor was typed in order to be entered in UNOS for matches with potential recipients. As UNOS matching is based on serological equivalents, even if we had obtained the A*24:53 result, we would not have reported it any differently in UNOS (i.e., we would have still reported A*24). Moreover, though we have the ability to obtain more information with NGS, time remains a limiting factor for initial HLA typing in cadaveric donors. Additionally, since matching in UNOS continues to be based on serological equivalents, all the advantages of NGS typing will be reserved for living donor and bone marrow transplants.

Human Immunology, 2019

Aim: The classical method for isolation of lymphocytes from peripheral blood is by use of Ficoll-... more Aim: The classical method for isolation of lymphocytes from peripheral blood is by use of Ficoll-Paque (F-Q) density gradient. This method is labor intensive. Isolated lymphocytes are often contaminated with Polymorphonuclear (PMN) and red blood cells (RBC) which requires additional methods for purification. The aim of our study was to determine if isolation of peripheral blood lymphocytes (PBL) using CPT tubes will shorten the isolation process, reduce the labor-intensive steps and yield a pure lymphocyte for testing. Methods: Equal volume of peripheral blood were isolated by the standard F-Q method and CPT tubes (BD Biosciences). The standard method of isolation by F-Q requires dilution, under layering and harvesting of lymphocytes. The CPT tube method requires pouring mixed blood into CPT tubes, spinning, pouring out cells above gel layer into another tube for wash step. The cells isolated by both methods were used in Flow and CDC crossmatches to determine the cell viability, cell count, and reproducibility of test results. Results: In 90% of samples, the number of mononuclear cells by CPT method were 12-100% higher than the F-Q method. The percentage of PMNs contamination was the same for both methods. In general, cell viability of cells isolated by both methods were above 90%. For older samples, cell isolated by the CPT method had a greater than 50% viability and lower background compared to the F-Q method. Results of CDC and flow crossmatch performed using cells isolated by both methods were similar and consistent with patient antibody history. The hands-on time for processing cells with CPT methods was 50% shorter than F-Q method. The CPT method eliminated cell harvesting step that is a major cause of hand injury. The cost of the F-Q and CPT method was the same. Conclusions: Isolation of lymphocytes using CPT tubes requires less processing time and better cell yield than standard F-Q separation method that is used by many HLA laboratories. Although the CPT tubes are more expensive than the regular isolation method, the total cost of isolation is the same due to the reduction in tech time spent in processing. Cells isolated by CPT method yields more viable cells when frozen and thawed for testing. Reduction in hand injury due to harvesting is another advantage to this new method.

Human Immunology, 2017

Aim: Determine if the COBAS Ò AmpliPrep/COBAS Ò TaqMan Ò System (COBAS Ò System) could be used to... more Aim: Determine if the COBAS Ò AmpliPrep/COBAS Ò TaqMan Ò System (COBAS Ò System) could be used to quantify viral loads of HIV and HBV in FFPE biopsies using as input sample purified RNA and DNA, respectively. Methods: Cell lines containing HIV (CRL-8993 producing multiple non-infective copies of the HIV genome), HBV (CRL-2235 containing the genome of the HBV) and negative (Jurkat Clone E6-1) were used. Formalin treated (fixed) and non-treated cells suspended in SPEX buffer were analyzed in COBAS Ò System. RNA from non-treated cells was purified using QIAamp RNA Blood Mini Kit (Qiagen). RNA or DNA of fixed cells and FFPE cells were purify in a Maxwell Ò 16 LEV (Promega) with the corresponding Maxwell Ò 16 LEV FFPE RNA or DNA extraction kit. RNA and DNA were diluted in SPEX buffer for analysis. Results: HIV cp/ml obtained in the COBAS Ò System with fixed CRL-8993 cells were 100 times lower than with non-treated cells. HIV cp/ml obtained with non-treated cells was similar to counts obtained with RNA from the same number of cells. Fixed cells RNA gave about half the cp/ml obtained by non-treated cells RNA. The drop was due to lower fixed cells RNA recovery, as adjusted values to cp/ng of RNA added showed no difference between fixed and non-treated cells. FFPE cells were prepared from dilution series of CRL-8993 into jurkat cells, maintaining a constant cellular density. The cp/ml and cp/lg of RNA obtained with RNA from FFPE curls of each cellular dilution showed high degree of correlation with the number of FFPE CRL-8993 cells. Quantitation of HBV was possible in CRL-2235 cells suspended in SPEX buffer. However, the same was not achieved in fixed cells. Quantitation of HBV in DNA from fixed and non-treated cells was very similar, with superimpose best fit equations. Conclusions: The COBAS Ò System can be used in the clinical laboratory to quantitate HIV and HBV in purified RNA and DNA from FFPE samples, respectively. There was no detectable loss of input RNA. This was not directly tested for DNA. It was revealed that the origin of nucleic acid was not a factor in the test as long as the appropriate purification method was applied. The methodology of this study could be extended to quantify HIV and HBV in RNA and DNA from fresh tissue biopsies. The procedure can be extended to other RNA and DNA viruses like HCV and CMV, respectively, for which kits are available.

Human Immunology, 2017

Aim: After a year of implementing HLA typing by NGS, we present our most relevant achievements an... more Aim: After a year of implementing HLA typing by NGS, we present our most relevant achievements and challenges. Methods: Class I (full HLA-A,-B, and-C genes) and HLA Class II (Exon 2 thru intron 3 of HLA-DR and HLA-DQ, and Exon 2 thru 3'UTR of HLA-DP) amplified with NXType TM Class I and Class II primer sets. Automated template preparation and chip loading were performed in the Ion Chef TM system and run on Ion S5 TM. Data was analyzed with TypeStream TM v1.1.0.11 software. Results: Having typed over 1000 samples by NGS, with 50% from bone marrow programs and 50% from solid organ programs, our workflow has changed significantly: Threefold increase in capacity with simultaneous decrease in turnaround time (TAT, on average 10 days) for high resolution typing. Our current capacity is up to 96 HLA typings in 3-4 days vs. minimum of 16 days for the same volume with SBT by Sanger. Resolution of most ambiguities (98.7%), minimizing the need for reflex testing.Repetitions due to technical failures and ambiguities decreased from 4.3% with SBT-Sanger to 0.3% with NGS. In spite of positive workflow changes, there remain challenges: Automation of library preparation has not been implemented.No merging of library preparation reagents into a single kit.Smaller runs increase the number of reads, leading to artifacts and high background generated by the software, interfering with analysis and limiting flexibility for STAT sample processing.For solid organ typings, generating high resolution results while reporting Serological Equivalents for UNOS, poses logistical difficulties.TypeStreamTM software not interfaced with Fusion, requiring continuous need to run SSOP for DSA analysis.Unresolved ambiguities (1.3%), due to incomplete exon coverage and phase ambiguities. Resolution may be obtained by increasing gene coverage, length of reads and with additional testing methods. Conclusions: We have confirmed that NGS is both a suitable and faster method to obtain high resolution HLA typing. We have seen a significant decrease in TAT along with a significant cost savings due to fewer repetitions, fewer reagents, and most significantly, reduced time from our most valuable asset, our technologists.

Human Immunology, 2017

We have established the Ion-torrent based Next Generation Sequencing (NGS) system in our clinical... more We have established the Ion-torrent based Next Generation Sequencing (NGS) system in our clinical laboratory. The chief use of the combination Ion-Chef TM and Ion-S5 is for high resolution HLA typing. However, the full sequencing capacity of the system is not always reached with the HLA typing workflow. In this study, we validated the practicality of combining library pools of different clinical tests into a single NGS run. Methods: HLA library pools of 24-32 patient samples were generated with NXType TM Ion-Torrent NGS for Class I (HLA-A,-B, and-C) and Class II (HLA-DR, HLA-DQ, and HLA-DP). These libraries were run alone and then separately combined with library pools of other clinical NGS-based assays. These included: (1) our lab-generated custom tests (APOL1 genotyping (single gene, three SNPs) and Cholestasis genotyping (15 genes, multiple SNPs and INDELS), and (2) commercial assays (16S-Metagenomics, Thermo Fisher Scientific, Inc.) and Lymphotrack, T and B cell clonality (Invivoscribe). All library pools were run on Ion-Chef TM and Ion-S5. HLA data was analyzed with TypeStream TM software v1.1.0.11. The concentration of HLA library pools was maintained at 100 pM. Importantly, the concentration of library pools of other tests were adjusted between 5 and 20 pM, depending on the test (number of genes and size of amplicons) and the number of samples in the pool. Exported data of APOL1, Cholestasis and 16S-Metagenomics were analyzed with Ion Reporter TM software. Lymphotrack data was exported and analyzed with Lymphotrack data analysis software. Results: The number of reads covering HLA genes were slightly reduced when runs were combined with libraries of other tests, however, the results of HLA library pools run alone and combined were 100% concordant. The coverage and read numbers obtained for the other tests were sufficient or exceeded the minimal required. Conclusions: To our knowledge, this is the first report showing that multiplexing different assays in a single run of Ion-S5 was achievable. We demonstrated that mixing sequencing samples of different genotyping tests in a single NGS run generated accurate results of each individual test. The findings of this works have a positive impact reducing turnaround times and sequencing costs.

Human Immunology, 2016

Aim We developed a custom TLDA to monitor changes of gene expression in FFPE renal and small bowe... more Aim We developed a custom TLDA to monitor changes of gene expression in FFPE renal and small bowel allografts. Here we carried out a parallel comparison of expression profiles of marker genes obtained with TaqMan® technology and with the automated qNPA system. Methods Kidney, small bowel and heart transplant FFPE biopsies with different levels of rejection were selected. 5–6 curls of 10 μm were cut from each biopsy for analysis in TLDA (47 markers including immune, inflammatory and apoptosis genes), and in HTG Edge System using the Immuno-Oncology Expression assay (HTGIOE, 26 markers), or HTG EdgeSeq Oncology Biomarker assay (HTGESOB, 2558 genes, format for NGS). Data was presented as differential expression. Results There are 10 overlapping markers between our TLDA and the HTGIOE assay. Analysis of 30 kidney biopsies with both technologies gave similar results for all 10 markers. Several markers were not detected in some samples analyzed with the HTG Edge System, mostly in normal donor and non-rejection samples. 11 unique markers of HTGIOE assay showed potential value for monitoring gene expression in kidney. 55 samples of heart biopsies were analyzed in TLDA and HTGIOE. Similar results were obtained with both technologies for all overlapping markers. 15 useful markers for monitoring gene expression in heart were found in HTGIOE assay. HTGIOE markers often did not express in non-rejection samples. We also analyzed 17 small bowel biopsies in HTGESOB, which contains 39 of the 47 markers in our TLDA (8 for SB, 17 for both kidney and small bowel, and 14 for kidney). 17.3% of the 2558 genes in the panel showed 2-fold increase expression relative to non-rejection. 15 markers in our TLDA (6 small bowel specific, 7 for both kidney and small bowel, and 2 kidney specific) were in the 5% of markers with the highest fold change. Conclusions We showed that expression analysis carried out with TLDA and qNPA are comparable. The qNPA technology developed by HTG Molecular Diagnostics, Inc. is a good alternative to assess expression in biopsies. We found that the HTGIOE could be an excellent automated tool to evaluate panels with low number of markers in a clinical lab. The HTGESOB allows evaluation of a large number of genes, but it includes a NGS step that makes it costlier and time consuming. It is a good exploratory tool for searching new markers.

Human Immunology, 2015

There are varying roles for facilitating, effector and regulatory CD4 helper cell subsets in soli... more There are varying roles for facilitating, effector and regulatory CD4 helper cell subsets in solid organ allograft maintenance and survival. Interplay between the relative levels may impact the character of the recipient’s anti-graft immune response. As such, efficient and accurate monitoring of their peripheral fluctuation can be a critical component of immune monitoring of transplant patients. In this study we developed a real-time PCR (RT-PCR) based TaqMan® algorithm to rapidly determine the proportions of Th1, Th2, Th17, and Treg cells in purified CD4 cells, using the expression markers TBX21, GATA3, RORC and FOXP3, respectively, with 18S as normalizing marker. The algorithm was tested with blood samples of 22 healthy donors and the expression results were compared and correlated with percentage of CD4 subsets obtained by intranuclear (FoxP3) and cytoplasmic (INF γ , IL4 and IL17A) staining flow cytometry. A strong correlation was found between expression of TBX21 and % of INF γ cells ( r = 0.60, p p r = 0.75, p r = 0.67, p γ /IL4; r = 0.73, p r = 0.57, p r = 0.50, p

British journal of pharmacology, 2014

The OX40-OX40L protein-protein interaction (PPI) is an important cell-surface signalling co-stimu... more The OX40-OX40L protein-protein interaction (PPI) is an important cell-surface signalling co-stimulatory regulator within the TNFR superfamily (TNFRSF) and a promising therapeutic target for immunomodulation. PPIs are difficult to modulate using small-molecules. Here, we describe the identification of a small-molecule OX40 modulator and confirm its partial agonist character. Cell-free screening assays were developed and used to identify OX40-OX40L inhibitors. Modified versions of this assay were used to elucidate the binding partner and the binding nature of active compounds. OX40-transfected sensor cells with NF-κB reporters were constructed and used to confirm and characterize activity and specificity. Immunomodulatory activity and partial agonist nature were further confirmed by ex vivo T-cell polarization assays. Several compounds that concentration-dependently affected OX40-OX40L were identified. Cell assays indicated that they were partial agonists with low micromolar potency a...

Pharmazie, 2012

As part of our ongoing effort to develop biohybrid devices for pancreatic islet transplantation, ... more As part of our ongoing effort to develop biohybrid devices for pancreatic islet transplantation, we are interested in establishing the feasibility of a localized immune-suppressive approach to avoid or minimize the undesirable side effects of existing systemic treatments. Since biohybrid devices can also incorporate biocompatible scaffold constructs to provide a support environment for the transplanted cells that enhances their engraftment and long-term function, we are particularly interested in an approach that would use the same three-dimensional construct, or part of the same construct, to also provide sustained release of therapeutic agents to modulate the inflammatory and immune responses locally. Within this framework, here, we report preliminary results obtained during the investigation of the suitability of organosilicone constructs for providing sustained localized drug release using small, matrix-type polydimethylsiloxane (PDMS) disks and dexamethasone as a model hydrophobic drug. Following a short burst, long-term steady sustained release was observed under in vitro conditions at levels of 0.1-0.5 g/day/disk with a profile in excellent agreement with that predicted by the Higuchi equation. To verify that therapeutic levels can be achieved, suppression of LPS-induced activation has been shown in THP-1 cells with disks that have been pre-soaked for up to 28 days. These preliminary results prove the feasibility of this approach where an integral part of the biomaterial construct used to enhance cell engraftment and long-term function also serves to provide sustained local drug release.

European Journal of Biochemistry, 1996

The axel gene encoding acetyl xylan esterase was isolated from an expression library of the filam... more The axel gene encoding acetyl xylan esterase was isolated from an expression library of the filamentous fungus Trichoderma reesei using antibodies raised against the purified enzyme. Apparently axel codes for the two forms, PI 7 and PI 6.8, of acetyl xylan esterase previously characterized. The axel encodes 302 amino acids including a signal sequence and a putative propeptide. The catalytic domain has no amino acid similarity with the reported acetyl xylan esterases but has a clear similarity, especially in the active site, with fungal cutinases which are serine esterases. Similarly to serine esterases, the axel product was inactivated with phenylmethylsulfonyl fluoride. At its C-terminus it carries a cellulose binding domain of fungal type, which is separated from the catalytic domain by a region rich in serine, glycine, threonine and proline. The binding domain can be separated from the catalytic domain by limited proteolysis without affecting the activity of the enzyme towards acetylated xylan, but abolishing its capability to bind cellulose.

European Journal of Biochemistry, 1996

Three cx-galactosidase genes, agll, ag12 and ag13, were isolated from a cDNA expression library o... more Three cx-galactosidase genes, agll, ag12 and ag13, were isolated from a cDNA expression library of Trichoderma reesei RutC-30 constructed in the yeast Saccharomyces cerevisiae by screening the library on plates containing the substrate 5-bromo-4-chloro-3-indolyl-aD -gnlactopyranoside. The genes ugll, ag12 and agl3 encode 444, 746 and 624 amino acids, respectively, including the signal sequences. The deduced amino acid sequences of AGLI and AGLIII showed similarity with the a-galactosidases of plant, animal, yeast and filamentous fungal origin classified into family 27 of glycosyl hydrolases whereas the deduced amino acid sequence of AGLII showed similarity with the bacterial a-galactosidases of family 36. The enzymes produced by yeast were analysed for enzymatic activity against different substrates. AGLI, AGLII and AGLIII were able to hydrolyse the synthetic substrate p-nitrophenyl-aD -galactopyranoside and the small galactose-containing oligosaccharides, melibiose and raffinose. They liberated galactose from polymeric galacto(g1uco)mannan with different efficiencies. The action of AGLI towards polymeric substrates was enhanced by the presence of the endo-l,4-/?-mannanase of T reesei. AGLII and AGLIII showed synergy in galacto(g1uco)mannan hydrolysis with the endo-l,4-/?-mannanase of 7: reesei and a B-mannosidase of Aspergillus niger. The calculated molecular mass and the hydrolytic properties of AGLI indicate that it corresponds to the a-galactosidase previously purified from 7: reesei.

Journal of Molecular Recognition, 2009

It is becoming increasingly clear that small molecules can often act as effective protein-protein... more It is becoming increasingly clear that small molecules can often act as effective protein-protein interaction (PPI) inhibitors, an area of increasing interest for its many possible therapeutic applications. We have identified several organic dyes and related small molecules that (i) concentration-dependently inhibit the important CD40-CD154 costimulatory interaction with activities in the low micromolar (microM) range, (ii) show selectivity toward this particular PPI, (iii) seem to bind on the surface of CD154, and (iv) concentration-dependently inhibit the CD154-induced B cell proliferation. They were identified through an iterative activity screening/structural similarity search procedure starting with suramin as lead, and the best smaller compounds, the main focus of the present work, achieved an almost 3-fold increase in ligand efficiency (DeltaG(0)/nonhydrogen atom = 0.8 kJ/N(nHa)) approaching the average of known promising small-molecule PPI inhibitors (approximately 1.0 kJ/N(nHa)). Since CD154 is a member of the tumor necrosis factor (TNF) superfamily of cell surface interaction molecules, inhibitory activities on the TNF-R1-TNF-alpha interactions were also determined to test for specificity, and the compounds selected here all showed more than 30-fold selectivity toward the CD40-CD154 interaction. Because of their easy availability in various structural scaffolds and because of their good protein-binding ability, often explored for tissue-specific staining and other purposes, such organic dyes can provide a valuable addition to the chemical space searched to identify small molecule PPI inhibitors in general.

Transplantation Pathology

Human Immunology, 2018

Applied and Environmental Microbiology, 1996

Applied and environmental microbiology, 1996

Applied and Environmental Microbiology, 1996

Human Immunology, 2019

Human Immunology, 2017

Human Immunology, 2016

Human Immunology, 2015

British journal of pharmacology, 2014

Pharmazie, 2012

European Journal of Biochemistry, 1996

Journal of Molecular Recognition, 2009