Lectin nanoparticle assays for detecting breast cancer-associated glycovariants of cancer antigen 15-3 (CA15-3) in human plasma

Abstract

Cancer antigen 15–3 (CA15-3) is widely utilized for monitoring metastatic breast cancer (BC). However, its utility for early detection of breast cancer is severely limited due to poor clinical sensitivity and specificity. The glycosylation of CA15-3 is known to be affected by BC, and therefore it might offer a way to construct CA15-3 glycovariant assays with improved cancer specificity. To this end, we performed lectin-based glycoprofiling of BC-associated CA15-3. CA15-3 expressed by a BC cell line was immobilized on microtitration wells using an anti-CA15-3 antibody. The glycosylation of the immobilized CA15-3 was then detected by using lectins coated onto europium (III)-doped nanoparticles (Eu +3 -NPs) and measuring the time-resolved fluorescence of Eu. Out of multiple lectin-Eu +3 -NP preparations, wheat germ agglutinin (WGA) and macrophage galactose-type lectin (MGL) -Eu 3+ -NPs bound to the BC cell line-dericed CA15-3 glycovariants (CA15-3 Lectin ). To evaluate the clinical performance of these two lectin-based assays, plasma samples from metastatic BC patients (n = 53) and healthy age-matched women (n = 20).Plasma CA15-3 Lectin measurements better distinguished metastatic BC patients from healthy controls than the conventional CA15-3 immunoassay. At 90% specificity, the clinical sensitivity of the assays was 66.0, 67.9 and 81.1% for the conventional CA15-3, CA15-3 MGL and CA15-3 WGA assays, respectively. Baseline CA15-3 MGL and CA15-3 WGA were correlated to conventional baseline CA15-3 levels (r = 0.68, p<0.001, r = 0.90, p>0.001, respectively). However, very low baseline CA15-3 MGL levels ≤ 5 U/mL were common in this metastatic breast cancer patient population.In conclusion, the new CA15-3 Lectin concept could considerably improve the clinical sensitivity of BC detection compared to the conventional CA15-3 immunoassays and should be validated further on a larger series of subjects with different cancer subtypes and stages.

A New Nanomaterial Based Biosensor for MUC1 Biomarker Detection in Early Diagnosis, Tumor Progression and Treatment of Cancer

“…If it is desired to measure at lower LOD values, experiments can be performed using a specific lectin for MUC1-C subgroups such as C-type lectin, galactose type lectin or other plant derived lectins instead of Concanavalin A. There are some studies about MUC1 detection based on other lectins [61][62][63]. Additionally, MUC1 aptamer and MUC1 antibodycan be used to provide higher analytical performances as found in the literature [64][65][66][67].…”

Nanoparticle-Aided Detection of Colorectal Cancer-Associated Glycoconjugates of Extracellular Vesicles in Human Serum

“…Extracellular vesicles (EVs) have been widely investigated in recent years, and emerging data suggest that cancer-derived EVs are strongly glycosylated, being rich in specific glycoconjugate [ 21 ]. In this study, we report on fluorescence europium chelate dyed-nanoparticles (Eu-NP)-based TRF assays to demonstrate the presence of specific proteins and glycans on the surface of EVs in CRC cell culture spend medium and further clinical evaluation of promising candidates in clinical serum samples–We have previously utilized the lectin-NP-based platform successfully to explore the glycosylation of serum glycoproteins CA125 (CA125 MGL ) and CA15-3 (CA15-3 WGA) in ovarian and breast cancer patients’ samples [ 31 , 32 ]. More recently, our group developed NP-based TRF assay for analysis and characterization of EVs and identification of disease-specific markers on the surface of patient-derived urinary EVs [ 30 , 33 ].…”

“…The Eu-NPs as bioconjugated reporter molecules have been described previously [ 47 ]. Amino groups of glycan-binding proteins (lectins), tetraspanins, and integrin monoclonal antibodies (mABs) were covalently bound to the activated carboxyl group of the Eu NPs according to the previously described procedure [ 28 , 32 ]. Briefly, N-hydroxysulfosuccinimide and N-(3-dimethylaminopropyl)-N′-ethylcarbodimide were incubated with 1e10 12 NPs in 50 mM 2-(N-morpholino) ethanesulfonic acid (MES) buffer (pH 6.1) at final concentrations of 10 and 0.75 mmol/L, respectively, at room temperature for 15 min.…”

Cell‐Membrane‐Display Nanotechnology

“…[40] The lectin-based assay is shown to be better than conventional immunoassay in using plasma samples to distinguish metastatic breast cancer patients from healthy controls. [41] boost immune reactions for disease prevention or treatment. Apart from escaping from clearance, the self-marker CD47 is also utilized to modulate host inflammatory response to synthetic polymeric surfaces.…”