Carrier Protein Research Articles

Transporters and drug secretion into human breastmilk.

Medication use is highly prevalent in breastfeeding persons, posing potential risks for drug exposure to nursing infants. Transporters in the lactating mammary gland carry pharmacological and toxicological significance, as they can mediate the active transfer of drugs and nutrients into breastmilk. In this narrative review, we searched and compiled current knowledge on the transport of drugs in the human mammary gland from literature indexed in PubMed (current as of 25 October 2024), and clinical evidence demonstrating active transport of drugs into milk is provided. In vitro and in vivo models of the mammary gland are outlined in brief and known drug transporters at the blood-milk barrier and their potential relevance to drug concentrations in milk are described in detail. Although clinical data show that membrane transporters mediate the transfer of multiple drugs into breastmilk, our ability to predict milk concentrations for these drugs is limited. Improving our understanding of the transporter biology and pharmacology in the mammary gland is crucial for developing models to predict drug concentrations in human milk, which will support clinicians and lactating individuals in making rational decisions to balance the benefits of breastfeeding and the risks of drug exposure to infants.

Membrane transport engineering for efficient yeast biomanufacturing.

Yeast strains have been widely recognized as useful cell factories for biomanufacturing. To improve production efficiency, their biosynthetic pathways and regulatory strategies have been continuously optimized. However, commercial production using yeasts is still limited by low product yield and high production cost. Accumulating evidences have demonstrated the importance of metabolite transport processes in addressing these challenges. Engineering yeast membrane transporters for transporting precursors, substrates, intermediates, products and toxic inhibitors has been successful. In addition, membrane properties are also important for metabolite production. Here we propose membrane transport engineering (MTE) to integrate manipulation of both membrane transporters and membrane properties. We emphasize that systematic optimization of both transporters and membrane lipid bilayers benefits production efficiency. We also envision the potential of artificial intelligence and automation process in MTE for economic and sustainable bioproduction using yeast cell factories.

Engineering an Escherichia coli surface display platform based on an autotransporter from Stenotrophomonas maltophilia: Autodisplay of enzymes with low to high molecular weight.

Surface display technology has garnered significant attention for preparing efficient whole cell catalysts, while reported carrier proteins still cannot meet the demand to display various passenger domains, especially for those with high molecular weight. This study demonstrates that the autotransporter of esterase Est7 (E7AT) from Stenotrophomonas maltophilia played a decisive role in its efficient surface display. Guided by the original signal peptide, the surface display ratio of Est7 was determined as 89.67% with the total enzymatic activity of 16.67U/mL, which was much higher than 4.60U/mL and 5.70U/mL for the signal peptides derived from pectolase B (pelB) and cholera toxin B (ctxB), respectively. Then, the E7AT unit was successfully developed to surface display proteins with varying molecular weight from 19.3kDa to 117.9kDa, showing a more effective autodisplay ability than adhesin involved in diffuse adherence (AIDA-I), ice nucleation proteins (InaK and InaP), and outer membrane proteins (lipoprotein/ompA, MltA-interacting protein A, and yiaT) systems. Additionally, a galactosidase (GAL) displayed by E7AT was employed to hydrolyze lactose, achieving a promising hydrolysis rate of 31.63% in 2h. The displayed GAL retained 63.24% and 41.41% activity in third and sixth batch, respectively, indicating the considerable potential of E7AT in developing efficient whole cell catalysts.

MXene-rGO nanocomposite based electrochemical immunosensor for detection of endosulfan - An organochlorine pesticide.

Endosulfan (Ed), a widely used organochlorine pesticide, is classified as a persistent organic pollutant (POP). Its long half-life, resistance to degradation, and bioaccumulation in the food chain contaminates soil, water, and air. Such widespread environmental damage triggers monitoring its levels for ensuring compliance with safety regulations and protecting public health. In the current work, Ed was chemically altered and coupled with a carrier protein to elicit an immunological response. The purified in-house generated antibodies against Ed (Ed-Ab) were characterized by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). MXene, a class of 2D inorganic compounds, is known to depict significant optoelectrical potential. Herein, we have synthesized a novel nanocomposite of MXene and reduced graphene oxide (rGO). For designing the MXene-rGO biosensor, Ed-Ab were combined with the nanocomposite post characterization by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDAX), X-ray photoelectron spectroscopy (XPS) and Fourier Transform infrared spectroscopy (FTIR). Using the differential pulse voltammetry (DPV), electrochemical parameters such as pH, temperature, scan rate and response time of the electrode were evaluated. The fabricated electro-immunosensor was employed for the detection of endosulfan wherein the limit of detection (LOD) for Ed was determined to be 0.497ppt with a linear range of 0.1ppt-1ppm. The composed electrode's working efficacy and sensitivity against similar cross-reactive pesticides was also determined. The MXene-rGO based nanocomposite depicted potential for determination of Ed traces in environmental samples.

Synthesis, Antibacterial Activity and In Silico Study of 1-(2-ethyl acetate)-2-styryl 5-nitroimidazole Derivatives

Background: For more than six decades, the use of metronidazole has been limited to anaerobic microorganisms. However, there are accounts of metronidazole derivatives exhibiting strong effectiveness against facultative anaerobic bacteria, suggesting that there may be another mechanism of action for metronidazole. Recent studies have shown that the enzyme FabH (β-ketoacyl-acyl carrier protein synthase III), responsible for the first step of fatty acid biosynthesis (FAB), is a promising target for nitroimidazole derivatives that can be used as an effective anti-infective agent. Objective: This study aimed to synthesize 1-(2-ethyl acetate)-2-styryl nitroimidazole derivatives and evaluate their in vitro and in silico antibacterial activity Methods: We synthesized 2-styryl 5-nitroimidazole derivatives by first condensing metronidazole with benzaldehydes and then carrying out an acetylation reaction. We evaluated the antimicrobial activity of the synthesized compounds against three Gram-positive bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus agalactiae) and three Gram-negative bacterial strains (Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) using a two-fold serial dilution MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay. Results: Compounds 2 and 4 exhibited the highest level of antibacterial effectiveness, with minimum inhibitory concentrations (MIC) of 1.56 μg/mL against S. agalactiae and 3.13 μg/mL against P. aeruginosa. Compounds 2 and 4 also exhibited potent activity against K. pneumonia, with an MIC value of 6.25 μg/mL and 12.5 μg/mL, respectively. Molecular docking studies revealed that both compounds have favorable hydrophobic and electrostatic interactions with conserved residues in the binding site of the E. coli β-Ketoacyl-acyl carrier protein synthase III (FabH) complex. Conclusion:: Acetylation of 2-styryl-5-nitroimidazoles improved both their biological activity and binding interaction with the target protein

Lipid metabolism during seed germination of Pistacia chinensis and its response to gibberellic acid

Lipid metabolism may play a critical role in fueling seed germination, but the knowledge of lipid metabolism during germination is still ambiguous. Here, we hypothesize that gibberellic acid (GA) promotes germination by means of enhancing lipid mobilization in Chinese pistachio (Pistacia chinensis Bunge), a species belonging to Anacardiaceae with high oil content in its seeds. A multi-omics approach has been applied to measure lipid mobilization during seed germination, and to identify the key regulators involved in GA-mediated lipid metabolism. The results indicated that GA contents increased, while IAA, ABA and JA contents decreased during seed germination. GA3 increased significantly in the two germination stages (i.e. imbibition and radicle protrusion), and it was more abundant than GA1 and GA4. In addition, the relative content of most lipids decreased during germination, and the differentially changed metabolites were significantly enriched in lipid metabolic pathways based on KEGG analysis. WGCNA indicated that GA3 was correlated with more genes in lipid metabolic pathways. Transcriptomic analysis further revealed that differentially expressed genes (DEGs) related to fatty acid biosynthesis, glycerolipid metabolism, glycerophospholipid metabolism and starch and sucrose metabolism were upregulated under GA3 application, such as the acetyl-CoA carboxylase biotin carboxyl carrier protein (ACCB), fatty acyl-ACP thioesterase B (FATB), diacylglycerol acyltransferase (DGAT) and DEFECTIVE IN ANTHER DEHISCENCE 1 (DAD1). Therefore, our study supports the hypothesis that GA promotes seed germination in P. chinensis by enhancing lipid mobilization. This study proposes a novel mechanism of lipid responses to exogenous GA, which contributes to a deep understanding of germination of oleaginous seeds.

Expanding the scope of copper artificial metalloenzymes: A potential fluorinase?

Biocatalysts for fluorination are rare, and thus of great interest for artificial enzyme design. Biohybrid catalysts including Cu-based DNAzymes and dinucleotide catalysts can catalyse enantioselective electrophilic fluorination of β-ketoesters. Here we report the investigation of Cu-based artificial metalloenzymes as catalysts for electrophilic fluorination reactions. A library of artificial copper proteins was prepared by bioconjugation of bidentate and tridentate nitrogen ligands to cysteine variants of the Sterol Carrier Protein 2 L (SCP-2 L) and subsequent addition of Cu(II) salts. The resulting copper proteins were screened for activity for the fluorination of β-ketoesters using Selectfluor. Under aqueous acidic conditions it was observed that the designed catalysts did not outcompete the uncatalysed background reaction. This work highlights that careful consideration of substrate reactivity and background reactions is needed when considering potential reactions for artificial metalloenzyme catalysis.

Antibody-secreting cell repertoires hold high-affinity anti-rocuronium specificities that can induce anaphylaxis in vivo.

Neuromuscular blocking agents (NMBA) are muscle relaxants used to assist mechanical ventilation but lead in 1/10,000 anesthesia to severe acute hypersensitivity reactions i.e., anaphylaxis. Incidences vary between types of NMBAs. Rocuronium, a widely used non-depolarizing aminosteroid NMBA, induces among the highest anaphylaxis rates. Rocuronium-induced anaphylaxis is proposed to rely on pre-existing rocuronium-binding antibodies, but no such antibodies have ever been identified. Identify rocuronium-specific antibody repertoires from plasma cells or plasmablasts of rocuronium-immunized mice, and determine the affinities, structures and anaphylactogenic potential of these antibodies for rocuronium. Herein, we engrafted rocuronium onto carrier proteins allowing immunization of mice against rocuronium, screening for rocuronium-specific antibody responses, and sorting of rocuronium-specific plasma cells using droplet microfluidics coupled to single-cell antibody gene (VH and VL) sequencing. The two different repertoires of >500 VH-VL pairs were oligoclonal, comprised of three major clonal families, and displayed convergence. Expressed as human IgG1, these antibodies demonstrated subnanomolar affinities for rocuronium with families either monospecific for rocuronium or cross-reactive only for closely-related NMBAs. Expressed as human IgE, they triggered human mast cell and basophil activation, and severe passive systemic anaphylaxis in mice humanized for the IgE receptor FcεRI. Co-crystal structures between rocuronium and antibody representatives of three different VH-VL families revealed distinct interaction modes with, however, the ammonium group involved systematically in the binding interface. This work identifies the epitopes of antibody reactivity to rocuronium, demonstrates anaphylactogenic potential of anti-rocuronium IgE and establishes the first mouse model of NMBA anaphylaxis.

Comprehensive analysis of the succinylome in Vero cells infected with peste des petits ruminants virus Nigeria 75/1 vaccine strain.

Peste des petits ruminants virus (PPRV) is currently the only member of the Morbillivirus caprinae species within the genus Morbillivirus of the family Paramyoxviridae. PPRV causes a highly contagious disease in small ruminants, especially goats and sheep. Succinylation is a newly identified and conserved modification and plays an important role in host cell response to pathogen infection. However, the extent and function of succinylation in Vero cells during PPRV infection remains unknown. In this study, a global profile of the succinylome in Vero cells infected with PPRV Nigeria 75/1 vaccine strain (PPRVvac) was performed by dimethylation labeling-based quantitative proteomics analysis. A total of 2633 succinylation sites derived from 823 proteins were quantified. The comparative analysis of differentially succinylated sites revealed that 228 down-regulated succinylation sites on 139 proteins and 44 up-regulated succinylation sites on 38 proteins were significantly modified in response to PPRVvac infection, seven succinylation motifs were identified. GO classification indicated that the differentially succinylated proteins (DSuPs) mainly participated in cellular respiration, biosynthetic process and transmembrane transporter activity. KEGG pathway analysis indicated that DSuPs were related to protein processing in the endoplasmic reticulum. Protein-protein interaction networks of the identified proteins provided further evidence that various ATP synthase subunits and carbon metabolism were modulated by succinylation, while the overlapped proteins between succinylation and acetylation are involved in glyoxylate and dicarboxylate metabolism. The findings of the present study provide the first report of the succinylome in Vero cells infected with PPRVvac and provided a foundation for investigating the role of succinylation alone and its overlap with acetylation in response to PPRVvac.

SLC29A1 and SLC29A2 are human nicotinamide cell membrane transporters

Nicotinamide (NAM), a main precursor of NAD+, is essential for cellular fuel respiration, energy production, and other cellular processes. Transporters for other precursors of NAD+ such as nicotinic acid and nicotinamide mononucleotide (NMN) have been identified, but the cellular transporter of nicotinamide has not been elucidated. Here, we demonstrate that equilibrative nucleoside transporter 1 and 2 (ENT1 and 2, encoded by SLC29A1 and 2) drive cellular nicotinamide uptake and establish nicotinamide metabolism homeostasis. In addition, ENT1/2 exhibits a strong capacity to change the cellular metabolite composition and the transcript, especially those related to nicotinamide. We further observe that ENT1/2 regulates cellular respiration and senescence, contributing by altering the NAD+ pool level and mitochondrial status. Changes to cellular respiration, mitochondrial status and senescence by ENT1/2 knockdown are reversed by NMN supplementation. Together, ENT1 and ENT2 act as both cellular nicotinamide-level keepers and nicotinamide biological regulators through their NAM transport functions.

Recent advances in the biosynthesis of polysaccharide-based antimicrobial glycoconjugate vaccines

Glycoconjugate vaccines are a vital category of effective and safe commercial vaccines that have significantly reduced the global prevalence of drug-resistant bacterial infections. These vaccines are synthesized by covalently linking bacterial polysaccharide antigens to a carrier protein. Given that they produce a stronger and longer-lasting immune response than pure polysaccharides that activate only B cells, glycoconjugate vaccines have become one of the most promising vaccine types. However, the chemical synthesis of glycoconjugate vaccines is complex, costly, and labor-intensive. Therefore, the efficient preparation of biosynthetic glycoconjugates using microbial cell factories has emerged as a highly desirable manufacturing alternative. This review focuses on advancements in the recombinant microbial biosynthesis of glycoconjugate vaccines and summarizes various strategies to optimize their production. It is based on three key aspects: the selection of oligosaccharyltransferase (OST), the use of different vaccine carrier proteins, and the enhancement of key concentrations in the uridine diphosphate (UDP)-sugar supply. Finally, the review highlights technical challenges and discusses future directions for the recombinant synthesis of glycoconjugate vaccines.

University students’ misconceptions of cellular structures and functions

ABSTRACT The goal of the present study was to assess biology students’ level of understanding and identify their misconceptions about cellular concepts. We conducted a questionnaire survey followed by interviews. Study participants (n = 620) were students enrolled in a biology degree programme. Data analysis showed that students had low knowledge of the basic characteristics of cells. Misconceptions were identified in all four core competencies of cell biology education. The most identified common misconceptions concerned the relationship between the structure and function of the cell membrane, nucleus and endoplasmic reticulum. Students have difficulty making the connection between cellular transport and its relationship to the structure of the plasma membrane. Students also struggled to make the connection between the different levels of organisation (macroscopic, microscopic and molecular). However, most students understood the concepts related to the chemical constituents of cells. Several potential factors are responsible for the appearance of misconceptions among the participants in this study, including levels, failure to use learning resources to visualise the various phenomena that take place in the cell, incomprehensible terms, and complexity of the processes and mechanisms of cell biology which, to understand them, require recourse to concepts taught in other disciplines such as physics and chemistry.

Порушення іонтранспортувальних процесів у сперматозоїдах неплідних чоловіків

During the study of the plasma membrane of spermatozoa and its organelles, a great variety of specifically localized ion channels, exchangers, and ATPases was discovered. Although the activity of ion channels and membrane transporters has been studied in detail, their involvement in the mechanisms leading to the dysfunction of male germ cells remains insufficiently elucidated. Numerous scientific works show that the absence of certain transport systems of the plasma membrane due to genetic mutations or their low activity leads to a decrease or loss of sperm motility, morphological changes that worsen the quality of sperm, and is the cause of male infertility. The review examines some ion transport systems that maintain resting membrane potential and ion homeostasis in spermatogenesis. It notes the possibility of using ion channels and membrane transporters as markers to establish the functionality of spermatozoa or as molecular targets for drugs in the treatment of male infertility.

Biosynthetic characterization of bacillibactin in thermophilic Bacillaceae and its potential forin vitro mutational synthesis.

Bacillibactin (BB) is a microbial siderophore produced by Bacillus species. BB is biosynthesized from 2,3-dihydroxybenzoic acid (2,3-DHB), Gly, and L-Thr by nonribosomal peptide synthetase (NRPS) enzymes DhbE, DhbB, and DhbF. The biosynthetic gene cluster (dhb) is also conserved in some strains of thermophilic genera, Geobacillus, Anoxybacillus and Parageobacillus. However, the production of BB from these thermophilic bacteria has not been characterized. Here, we report in vivo and in vitro characterization of BB biosynthesis in Parageobacillus sp. KH3-4 which grows at 65°C. We confirmed BB production in this thermophilic bacterium and the gene cluster active. In vitro enzymatic analysis revealed that 4'-phosphopantetheinyltransferase (PPTase) encoded in the same gene cluster is responsible for the post-translational maturation of carrier proteins. DhbE and DhbF showed substrate preference to 2,3-DHB and Gly and L-Thr, respectively, consistent with the chemical structure of BB. With the purified enzymes, we successfully reconstituted the NRPS assembly line in vitro. In addition, using chemically synthesized acyl-N-acetylcysteamine substrate analogues, BB analogues possessing methylbenzoyl groups instead of 2,3-DHB were detected. This study provides a new insight into secondary metabolism in thermophiles, and it expands the temperature limitation of NRPS enzymes.

Evaluation of the Role of PnuC Gene in Enhancing Nicotinamide Mononucleotide Synthesis.

The PnuC gene plays a crucial role in the complex processes related to the absorption and synthesis of the nicotinamide mononucleotide (NMN) precursor. NMN, a nicotinamide adenine dinucleotide (NAD+) precursor, is important for cellular energy metabolism, DNA repair, and antiaging. This study focuses on elucidating the precursor absorption mechanism and the specific function of the PnuC gene in encoding membrane transport proteins, as well as its impact on the regulation and dynamics of NMN within the cell. This understanding aims to provide insights into its potential effects on metabolic balance, illustrated through two NAD+ biosynthesis pathways based on renewable and readily available cytoplasmic resources, assessing the potential of PnuC gene expression in clarifying complex interactions within regulation mechanisms. Enhanced expression analysis of the PnuC gene has initiated discussions on its potential applications in treating aging-related diseases and dysfunctions, contributing to cellular health maintenance.

Potential roles of mitochondrial carrier proteins in plant responses to abiotic stress.

The transport of metabolites across the inner mitochondrial membrane (IMM) is crucial for maintaining energy balance and efficient distribution of metabolic intermediates between cellular compartments. Under abiotic stress, mitochondrial function becomes particularly critical, activating complex signaling pathways essential for plant stress responses. These pathways modulate stress-responsive gene expression, influencing key physiological processes such as cell respiration and senescence, helping plants adapt to stress. Recent studies have emphasized the importance of finely tuned regulation of mitochondrial metabolite transport through the IMM, particularly under stress conditions, to optimize plant survival and resilience. This review summarizes the current knowledge on the possible roles of mitochondrial transport proteins and their contributions to plant adaptation under abiotic stress.

Urobilin Derived from Bilirubin Bioconversion Binds Albumin and May Interfere with Bilirubin Interacting with Albumin: Implications for Disease Pathology

Background/Objectives: Bilirubin is a hydrophobic molecule that binds the carrier protein albumin for transport through systemic circulation. Bilirubin is cleared from the body through the liver and excreted into the intestines, where the microbiota modifies the chemical structure, forming urobilin, which can be reabsorbed into circulation by the hepatic portal vein. Urobilin has no known function. It is also unknown whether urobilin binds albumin for transport in circulation. We hypothesized that because of the likeness of their chemical structures, urobilin would also bind albumin like bilirubin does. Methods: First, we used in silico docking to predict if urobilin would bind to albumin and compared it to the bilirubin binding sites. To test this binding in vitro, we applied bilirubin’s fluorescent property, which occurs when it is bound to a protein, including albumin, and exposed to light. We also used this method to determine if urobilin could exhibit autofluorescence when protein bound. Results: We found that bilirubin was predicted to bind albumin at amino acids E208, K212, D237, and K240 through hydrogen bonds. However, urobilin was predicted to bind albumin using different hydrogen bonds at amino acids H67, K240, and E252. We found that urobilin has a fluorescent property that can be quantified when bound to albumin. We performed a concentration response for urobilin–albumin fluorescent binding and observed a direct relationship between the urobilin level and the fluorescence intensity. Conclusions: The in silico docking analysis and autofluorescence results demonstrate that urobilin binds to albumin and might compete with bilirubin. This is the first study to identify a urobilin-binding protein and the important aspects of its physiological function and transport in circulation.

Elucidation of interface interactions between a dehydratase domain and an acyl carrier protein in cremimycin polyketide synthase.

Modular polyketide synthases (PKSs) are multi-domain enzymes involved in the biosynthesis of polyketide natural products. The dehydratase (DH) domain catalyzes the dehydration of the β-hydroxyacyl unit attached to the acyl carrier protein (ACP) domain in modular PKS. Although the DH domain likely recognizes the cognate ACP domain during the dehydration reaction, the molecular basis of DH-ACP interactions remains elusive. In this study, we conducted cross-linking analysis using a pantetheine-type probe for investigating the ACP recognition of a fusion-DH protein generated from a split-DH domain of cremimycin PKS. Based on the AlphaFold 3-predicted model structure of the fusion-DH-ACP complex, DH-ACP interface residues were identified and validated by mutational analysis. Our findings provide the first detailed insights into domain-domain interactions between DH and ACP in modular PKSs.

The Salmonella Paratyphi A O-Antigen Glycoconjugate Vaccine Is Able to Induce Antibodies with Bactericidal Activity Against a Panel of Clinical Isolates

Background: Typhoid and paratyphoid fevers represent a global health burden, especially in Southern Asia, exacerbated by the increase in antimicrobial resistance. While vaccines against Salmonella Typhi have been successfully introduced, a vaccine against S. Paratyphi A is not available, yet. Efforts to develop an effective vaccine targeting both Salmonella serovars are currently ongoing. GVGH is developing a bivalent vaccine constituted by the Vi-CRM197 typhoid conjugate vaccine (TCV), and the Salmonella Paratyphi A O-antigen (O:2), also conjugated to the CRM197 carrier protein (O:2-CRM197). In this work we have characterized a panel of S. Paratyphi A clinical isolates from endemic regions, differing in terms of their O:2 structural features. Methods: Rabbits were immunized with the S. Paratyphi A component of the vaccine candidate and the resulting sera were tested for their ability to bind and kill the isolates using flow cytometry and luminescence-based serum bactericidal assay (L-SBA). Results: The O:2-CRM197 glycoconjugate induced a functional immune response in rabbits, effectively binding and killing a diverse panel of clinical isolates. The sera demonstrated bactericidal activity independent of the O:2 structural variations, including differences in O-acetylation and glucosylation levels. Additionally, the study found that the O:2-CRM197 conjugate’s adsorption to Alhydrogel did not significantly impact its immunogenicity or bactericidal efficacy. Conclusions: The O:2-CRM197 component of the bivalent vaccine candidate shows promise in providing broad protection against S. Paratyphi A isolates, regardless of their O-antigen structural variations. The ongoing clinical studies on human sera are expected to confirm these results.

MUC1 and glycan probing of CA19-9 captured biomarkers from cyst fluids and serum provides enhanced recognition of ovarian cancer

Glycosylation changes of circulating proteins carrying the CA19-9 antigen may offer new targets for detection methods to be explored for the diagnosis of epithelial ovarian cancer (EOC). Search for assay designs for targets initially captured by a CA19-9 antigen reactive antibody from human body fluids by probing with fluorescent nanoparticles coated with lectins or antibodies to known EOC associated proteins. CA19-9 antigens were immobilized from ascites fluids, ovarian cyst fluids or serum samples using monoclonal antibody C192 followed by probing of carrier proteins using anti-MUC16, anti-MUC1 and, anti STn antibodies and seven lectins, all separately coated on nanoparticles. Compared to reference CA19-9 and CA125 immunoassays, nanoparticle aided detection using MUC16, Ma695 and STn antibodies and lectin WGA provided, both separately and combined, improved discrimination of EOC and borderline cancers from benign samples when applied to 60 cyst fluid specimens. When applied to a panel of 44 serum samples (EOC N = 24, healthy and benign samples N = 20) two assays, CA19-9Ma695 and CA19-9MUC1, stood out with equally superior separations (p-values < 10–8) of the two groups compared to conventional CA19-9 immunoassay (p-value 0.03).Eu+3 -NP based CA19-9MUC16, CA19-9Ma695, CA19-9STn and CA19-9WGA show promise for improved EOC detection when applied to ascites & cyst fluids. When applied to circulation-derived samples, the two MUC1 based assays, CA19-9Ma695 and CA19-9Ma552 outperformed other assay constructs. Our results call for further validation in larger EOC cohorts preferentially with early stage ovarian cancers and all major histotypes against commonly occurring benign conditions.