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[Full text] EDTA-K2 Improves the Detection Sensitivity of SARS-CoV-2 IgM and IgG A | IJN – Dove Medical Press

Introduction

Currently, the widespread threat to human health from SARS-CoV-2 urgently requires the development of fast and accurate analytical methods for early diagnosis. At present, pathogen detection and nucleic acid testing are commonly used to identify COVID-19 in the clinic. In addition, infected patients are mostly positive for SARS-CoV-2-specific IgM antibodies after seven to eight days from disease onset and their IgG titers during the recovery period, are more than four times higher than those in the acute phase,1 indicating that serological detection of specific antibodies is crucial for the rapid identification of SARS-CoV-2. Specifically, this study confirmed that the gold immunochromatographic assay (GICA) could serve as a rapid diagnostic test for RT-PCR-negative highly suspected patients and screening of SARS-CoV-2 carries in limited-resource countries.2,3 Recently, GICA has been widely used to detect antibodies, because the test trips based on colloidal gold particles were fast, convenient, and require minimal equipment. Therefore, we feel that this method is a good candidate for the large scale, rapid screening, of both symptomatic and asymptomatic COVID-19 carriers and the screening of healthy workers following isolation.4

Early in 2014, a study found that different additives in the blood collection tubes, were able to influenced the detection of alcohol concentration.5 Moreover, a study found that DNA samples extracted from blood using two different anticoagulants (heparin tubes and EDTA-K2), gave similar results, especially for PCR based applications.6 These studies revealed that different additives in the vacuum blood collection tubes, showed clear confounding effects clinically. However, it is not clear whether these vacuum blood collection tubes with different additives affect the detection of SARS-CoV-2 IgM and IgG antibodies. Therefore, this study investigated the impact of different vacuum blood collection tubes, when detecting SARS-CoV-2 IgM and IgG antibodies in the plasma and serum using GICA. These results are expected to provide a theoretical basis for the future accurate detection of SARS-CoV-2 IgM and IgG antibodies.

A total of 112 patients diagnosed as having COVID-19 by nucleic acid testing were enrolled from the Respiratory Department of Hankou Hospital of Wuhan, China, from March 6 to March 18, 2020. Nine of the 112 patients had their samples collected into four different types of collection tube. Among the patients, seven of the nine patients diagnosed with COVID-19 were used to quantify the concentration of SARS-CoV-2 IgM and IgG antibodies. Thirty-two of the 112 patients were used to detect SARS-CoV-2 IgM and IgG antibodies using EDTA-K2 as plasma and whole blood anticoagulants. Next, 200 healthy control subjects diagnosed as noninfectious for COVID-19 based on nucleic acid testing, were enrolled from the Department of Clinical Laboratory Medicine of Nanfang Hospital of Guangdong, China, from April 4 to April 5, 2020. Serum and plasma specimens were isolated from 2 mL of blood and placed into four different types of vacuum blood collection tube, which contained different additional reagents, such that the serum was collected into tubes with coagulants present and plasma with different anticoagulants (EDTA-K2, sodium citrate or lithium heparin).

IgG and IgM specific antibodies were detected in blood samples using recombinant coronavirus N protein labeled, and the results were seen using the naked eye. The new coronavirus antibody detection kit (colloidal gold based) and the colloidal gold solution were developed by Innotek (Tangshan) Biotechnology, and the chemiluminescence detection reagents were developed by Mairui Biotechnology Co., Ltd (Guangdong, China). The vacuum blood collection tubes with coagulants and EDTA-K2 were produced by Zhiyuan Biotechnology, Co., Ltd (Guangdong and Hubei, China), and those with sodium citrate and lithium heparin were from Jinxing Biotechnology Co., Ltd (Guangdong and Hubei, China). Blood samples from healthy subjects were collected into EDTA-K2 coated test tubes, and the samples from COVID-19 patients were collected into the test tubes as stated above. They were centrifuged at 1000g at 4C for 15 min and the serum and plasma collected. Finally, the samples were subjected to the GICA, or tested on an ARCHITECT i2000 (Mairui Biotechnology, Shenzhen, China) for the detection of IgM and IgG as specified by the manufacturer.

Immunocapture methodology was used here to detect the SARS-CoV-2 IgM and IgG antibodies. The nitrocellulose membrane was coated with mouse anti-human IgM monoclonal antibodies, mouse anti-human IgG monoclonal antibodies, and goat anti-mouse IgG antibodies. Colloidal gold-labeled recombinant coronavirus antigen and mouse IgG antibodies were then used as a tracer. When the sample was added to the well in the presence of the IgM or IgG detection reagent, it is able to combine with the corresponding antigen to form a complex and then the coated mouse anti-human antibodies will react with this complex, forming a purple-red band, as detected on the card. The colloidal gold-labeled mouse IgG antibodies will form a purple-red band with the goat anti-mouse IgG antibodies.

Here, our aim was to determine the effect of different additives in the collection test tubes on their ability to chelate the colloidal gold. The size of the particle was determined by a Zetasizer Nano ZS90 analysis system (Malvern Instruments, UK, Zetasizer version 7.12). A size polydispersity Index (PDI), for number particle size distribution (PSD), with the x-axis showing the distribution of estimated particle diameter (nm) and the y-axis showing the relative percentage, was created.

Data were analyzed using origin (version 2018), IBM SPSS Statistical software for Windows, version 22.0 (IBM Corporation Armonk, NY, USA) and GraphPad software (GraphPad Software, Inc., La Jolla, CA, USA). Data are presented as the mean standard deviation or median (interquartile range), unless otherwise indicated. The results from the concentration of SARS-CoV-2 antibodies detected by chemiluminescence assay (CA) were assessed by one-way analysis of variance followed by Student-Newman-Keuls test. A two-tailed probability (P) value of < 0.05 was considered statistically significant.

We collected serum and plasma with EDTA-K2 from 112 patients with COVID-19 to detect SARS-CoV-2 IgM and IgG antibodies using GICA. The vacuum collection tubes with coagulants and a separator were used to collect serum. Our results revealed that EDTA-K2 anticoagulated plasma had a higher positive detection rate for detecting SARS-CoV-2 IgM antibodies when compared to the serum group. Specifically, seventeen patients were found to be weakly positive, with an additional fifteen patients testing positive but were negative based on serum testing. Five patients were weakly positive, thirteen patients were positive and three patients were strongly positive, but very weak positive using serum testing. Twenty-one patients were detected as positive and two patient was strongly positive but weakly positive based on serum testing. Similarly, for IgG antibody testing, forty samples were interpreted as strongly positive but were detected as only positive when serum tested (Table 1).

Table 1 The Number of SARS-CoV-2 IgG and IgM Antibodies Detected in 112 Patients with Serum and Anticoagulated Plasma with EDTA-K2

In addition, we analyzed the consistency of the results in detecting SARS-CoV-2 IgM and IgG antibodies when comparing serum and EDTA-K2 anticoagulated plasma. We classified very weakly positive, weakly positive and strongly positive, as positive, and kappa values were used to evaluate the consistency of the positive and negative groups. We found the total coincidence rate of IgG antibodies were all 100% in these two groups (weighted kappa value = 1.0), while the kappa value for the IgM antibody was 0.409 between the two groups (Table 2). Taken together, these findings demonstrated that EDTA-K2 increased the positive detection rate of SARS-CoV-2 IgM antibodies.

Table 2 Analysis of the Consistency of Detecting SARS-CoV-2 IgG and IgM Antibodies in 112 Patients with Serum and Anticoagulated Plasma with EDTA-K2

Based on our hypothesis above, in order to investigate the impact of different types of vacuum collection tubes on the detection of SARS-CoV-2 antibodies, we collected blood specimens from nine COVID-19 patients with four different vacuum blood collection tubes (serum and anticoagulated plasma with lithium heparin, sodium citrate and EDTA-K2). Then, SARS-CoV-2 IgM and IgG antibodies were detected by GICA. Our results showed that EDTA-K2 anticoagulated plasma had a higher positive detection rate for IgM antibodies, with a darker color being present on the IgG card. However, there were no significant differences among the serum and anticoagulated plasma, when lithium heparin or sodium citrate were used (Figure 1).

Figure 1 Original picture on detected SARS-CoV-2 IgM and IgG antibodies in nine patients with different vacuum collection tubes by GICA. P: plasma (EDTA-K2); N: plasma (sodium citrate); L: plasma (lithium heparin); S: serum (coagulants).

In addition, we aimed to further verify that this highly positive effect was attributable to EDTA-K2. Therefore, thirty-two of the 112 patients with COVID-19 had their plasma collected into EDTA-K2 tubes and the whole blood from these patients was tested. Our data showed that there was no difference in the ability to detect SARS-CoV-2 IgM or IgG antibodies when both the anticoagulated blood plasma or whole blood with EDTA-K2 was used (Table 3 and Supplemental Figure 1). These results strongly demonstrated that anticoagulated blood plasma in the presence of EDTA-K2 improved the detection rates of SARS-CoV-2 IgM and IgG antibodies, and this was likely to be due to EDTA-K2, rather than the use of blood serum, plasma, or whole blood. Hence, these findings indicated a marked improvement when EDTA-K2 was used in the ability to enhance the detection of SARS-CoV-2 IgM antibodies.

Table 3 Percentage of SARS-CoV-2 Antibodies Detected in EDTA-K2 Anticoagulated Plasma and Whole Blood by GICA

To explore the ability of EDTA-K2 to improve the detection sensitivity of SARS-CoV-2 IgM and IgG antibodies, four different kinds of blood specimens from seven COVID-19 patients with four different vacuum blood collection tubes (serum and anticoagulated plasma with lithium heparin, sodium citrate and EDTA-K2), were then subjected to the GICA and CA, respectively. The latter assay has a higher sensitivity than the former.7 Our results revealed that when CA was used to detect SARS-CoV-2 IgM and IgG antibodies, the presence of different anticoagulants had no effect on the measurements for serum or plasma (Figure 2A and B). However, when the same samples were detected by GICA, the results became significantly variable (Figure 1). Compared with other anticoagulated plasma and serum specimens, the results using EDTA-K2 anticoagulated plasma detected by GICA were more consistent with those results using CA. For example, in patients from 36 to 39, the concentration of IgM antibodies was found to be 2.710.03, 2.840.09, 1.150.08, and 5.240.11 Au/mL respectively when using CA (the reference interval was 0.01.0 Au/mL). These patients would be expected to test positive or strongly positive using the GICA. However, only the results from plasma treated with EDTA-K2 were consistent with those from the CA. The other anticoagulated plasma samples were judged as negative or weakly positive by GICA (Figure 1 and Table 4).

Table 4 The Detection Results of SARS-CoV-2 IgM and IgG Antibodies in seven Patients by Colloidal Gold Assay and Chemiluminescence Assay

Figure 2 The detection results of SARS-CoV-2 IgM and IgG antibodies in seven patients by CA. (A). The detection results of SARS-CoV-2 IgM antibodies in seven patients. (B). The detection results of SARS-CoV-2 IgG antibodies in seven patients. Reference interval of IgM was 0.01.0 Au/mL; IgG was 0.010.0 Au/mL. #P>0.05 by one-way analysis of variance analysis.

We found that the anticoagulant EDTA-K2, increased detection sensitivity of the GICA. Next, we determined whether it induced the false positive for the detection of SARS-CoV-2 specific antibodies. We set up a blank group containing simply diluent and EDTA-K2. Surprisingly, our results showed that there was no visible purple-red color on the test line. Additionally, we collected anticoagulated plasma with EDTA-K2 from 200 healthy control patients with no COVID-19 infection, and all 200 healthy individuals were negative for both SARS-CoV-2 IgM and IgG antibodies, providing evidence that EDTA-K2 did not increase the false positive rate.

EDTA-K2 is commonly used in blood testing, as it can chelate calcium and reduce the anticoagulation reaction, However, it is unknown whether EDTA-K2 can also chelate colloidal gold and participate in antigen-antibody reactions. Therefore, we set up four groups: the control group was treated with the colloidal gold solution and coagulants, the test group was treated with EDTA-K2, sodium citrate or lithium heparin and colloidal gold solution. Next, Malvern apparatus was used to evaluate the particle size in the four groups. As expected, our results showed that there were two peaks in the EDTA-K2 group, but only one peak in other three groups.

Specifically, the particle sizes were 47.58 1.67 nm (PDI = 0.247 0.02), 49.62 2.20 nm (PDI = 0.250 0.04) and 48.27 2.65 nm (PDI = 0.209 0.07) in the Au, Au+sodium citrate and Au+lithium heparin groups respectively. However, for the Au+EDTA-K2 group, the particle size of first peak was 164 30 nm, which was larger than in the other three groups by approximately four times, and the second peak was 5399 143 nm (PDI = 0.275 0.03) (Figure 3A). Next, they were observed under a confocal laser microscope. We found that larger particle sizes in the test group with EDTA-K2 (Figure 3E), but no significant differences were seen between the Au group and the test groups containing lithium heparin, or sodium citrate (Figure 3BD). These results provided solid evidence in support of our hypothesis that EDTA-K2 could amplify the positive signal by chelating colloidal gold.

Figure 3 The particle sizes of colloidal gold were detected by laser diffraction (Malvern) and confocal laser microscope, and expressed as mean (SD) in four groups with five independent experiments, each performed in triplicate. (A) Particle sizes were 47.581.67 nm, 49.622.20 nm and 48.272.65 nm in the group of Au, Au+sodium citrate and Au+lithium heparin, respectively, but for the Au+EDTA-K2 group, the particle sizes of first peak was16430 nm, and the second peak was 5399143 nm. (BE) Four groups of Au, Au+lithium heparin, Au+sodium citrate and Au+EDTA-K2 were observed under the microscope with oil lens, respectively. Arrows: aggregated colloidal gold particles.

The incubation period of various SARS-CoV-2 infections in humans, is from one to fourteen days, and its transmission channels are diverse. In addition to the main routes of transmission through respiratory droplets and contact, it can also be transmitted through aerosols, the digestive tract, and vertical transmission.8 It has a high infection rate due to its multiple channels of transmission and strong infectivity. Therefore, early recognition of suspected patients, and identification of those with severe illness and the early isolation of those with clinical symptoms, are essential for disease control and prevention. At present, the nucleic acid SARS-CoV-2 RT-PCR tests have become the gold standard assay for the diagnosis of COVID-19 with a high sensitivity and specificity.9 However, this method is a relatively time-consuming and complicated and therefore, is not conducive to the screening of large numbers of samples.

GICA based on colloidal gold particles is widely used to screen for antibodies in the blood against SARS-CoV-2. It has the advantages of convenient, fast detection, low cost and suitable for rapid screening of large numbers of samples, the results are intuitive and there is no requirement for specialized equipment, etc.1012 Early in 2011, a study confirmed that GICA could be combine with the enrichment technique of immunomagnetic nanoparticles, to detect Escherichia coli O157: H7 with speed and sensitivity,13 this study provide powerful evidence for the conclusion that EDTA-K2 could chelate colloidal gold particles and increase the sensitivity of detection on SARS-CoV-2 IgM and IgG antibodies. However, some limitations exist for these assays, such as individual differences in the interpretation of results, which may lead to inconsistent results. In addition, these assays could be affected by several factors, including pH, where it was shown to influence the diagnosis of syphilis using GICA.14 Other studies reported a matrix effect when looking at five kinds of meat, using a colloidal GICA to detect sulfamethazine.15

EDTA-K2 has anticoagulant effects because it can chelate large amounts of calcium ions in the blood.16 Therefore, EDTA-K2 is commonly applied to blood samples as an anticoagulant in the clinical setting, but it can lead to the aggregation of platelets in the blood of individual patients, leading to pseudothrombocytopenia.17 EDTA-K2 has been found to influence the test results of Ca, Mg and Fe compared with the collection tubes excluding anticoagulant.18 Others have reported that EDTA-K2 can impact on the extent of detection of coagulation by PLT.19 However, researchers have found that EDTA-K2 no significant effects on the detection of divalent cation chelators containing anticoagulant detected ITP platelet specific antibodies (GP IIb/IIIa and GPI b) using multi-antigen printing immunoassays.20 Whether EDTA-K2 could affect the detection of antibodies based on GICA remains unknown. In this study, we confirmed that EDTA-K2 could improve the detection sensitivity of anti-SARS-CoV-2 specific antibodies. Previous study found that colloidal selenium melamine test strips could rapidly detect colloidal selenium melamine test strips in contaminated milk products or animal feed, with high sensitivity.21 This finding provided powerful evidence for our results that the method of EDTA-K2 based on colloidal gold could be used in the detection of SARS-CoV-2. Therefore, we concluded that there is a significant effect of EDTA-K2 on colloidal gold, but these mechanisms remain largely unknown, and more studies are needed to clarify this phenomenon.

IgM antibodies are rapidly produced in patients in response to acute infections with pathogens, therefore, it is important to accurately detect IgM antibodies at the time of initial diagnosis. In addition, IgM antibody tilters are important for monitoring recovering and relapsed patients. We found that EDTA-K2 could increase the positive detection rate of SARS-CoV-2 IgM antibodies when using GICAs. In this study, we designed a series of experiments to determine how EDTA-K2 affected the detection of anti-SARS-CoV-2 antibodies. We used confocal laser microscope and a Zetasizer Nano ZS90 analysis system to evaluate the particle sizes of our colloidal gold. The results revealed that EDTA-K2 could amplify the positive signal by chelating the colloidal gold. This suggested that, even if there was only a small amount of antibody in EDTA-K2 anticoagulated plasma, the color of the T line was darker. A deeper understanding of the reaction principle suggested that EDTA-K2 could amplify a weak signal by chelating colloidal gold and this can explain why the SARS-CoV-2 IgM antibodies could not be detected in other anticoagulated blood specimens such as sodium citrate and lithium heparin but could work in EDTA-K2 anticoagulated plasma.

Although its sensitivity is not as high as CA, GICA can easily be used to screen large numbers of samples and is suitable for use in primary hospitals. Currently, with the new coronaviruses spreading worldwide,22,23 it is urgent to improve the sensitivity and speed of detection of SARS-CoV-2 IgM and IgG antibodies. These are major priorities to help curb the spread of the virus, so it is important to ensure that any diagnosis is made promptly and accurately.

Overall, the results of this study provided solid evidence for the use of EDTA-K2 to improve the sensitivity of detection of SARS-CoV-2 IgM and IgG antibodies. However, it should be noted that when results between serum and anticoagulated plasma were inconsistent, we recommend that multiple methods should be used to confirm results, such as nucleic acid detection, CA, ELISA, and imaging examinations, combined with the patients clinical symptoms. These findings provided new insights for improving the detection of SARS-CoV-2 by GICA. However, several problems need to be addressed. First, the samples we used were from patients during the middle and late stages of treatment and rehabilitation. There 112 patients with COVID-19 and 200 healthy people involved in the project, but only seven patients with COVID-19 were enrolled to detect four different tubes. Since there are few patients infection SARS-CoV-2 in China, it is difficult to collect samples from the early stage of infection for verification, which is the limitation of this study. Second, other effects influencing the detection of COVID-19 by GICA remains unclear. Third, only one commercial kit was used for comparison in this study, and whether the pore size of the chromatography membrane had an effect on the test results remains largely unknown.

GICA and equipment-dependent CA were usually used to detect antibodies in clinic, GICA is widely used to detect antibodies for the advantages of convenient, fast, low cost, suitable for screening large sample and require minimal equipment. In this study, we found that EDTA-K2 anticoagulated plasma has higher positive rate than other anticoagulated plasma (sodium citrate and lithium heparin) or serum, especially, the results of EDTA-K2 anticoagulated plasma detected by GICA was high consistent with CA results. Further study shown that EDTA-K2 could amplify positive signal by chelating colloidal gold. Therefore, we suggested that EDTA-K2 anticoagulated plasma maybe more suitable for the detection of SARS-CoV-2 antibodies. However, more positive samples with SARS-CoV-2 were needed to further verify this results.

This study was approved by the Hankou Hospital Ethics Committee (No. HKYY-2020-028), and according to the Ethics Committee review, patients informed consent was not required. It is noteworthy that these specimens taken were part of routine hospital procedure, rather than being taken specifically for this study. There was no additional manipulation or injury performed on the participants. Everything was done to ensure that the patients personal information was maintained under strict privacy. This study complied with the Declaration of Helsinki.

This work was supported by funds from the National Natural Science Foundation of China (81601819), the Outstanding Youths Development Scheme of Nanfang Hospital, Southern Medical University (2016J013), the Medical Science and Technology Research Foundation of Guangdong Province (A2016280) and Funds for prevention and control of major infectious diseases from China government in 2020.

The authors report no conflicts of interest in this work.

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9. Corman VM, Landt O, Kaiser M, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25(3):2000045. doi:10.2807/1560-7917.ES.2020.25.3.2000045

10. Ray M, Achary KG, Nayak S, et al. Development of a colloidal gold strip-based immunochromatographic assay for rapid detection of Fusarium oxysporum in ginger. J Sci Food Agric. 2019;99(14):61556166. doi:10.1002/jsfa.9859

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[Full text] EDTA-K2 Improves the Detection Sensitivity of SARS-CoV-2 IgM and IgG A | IJN - Dove Medical Press

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Clene Nanomedicine Aims to be the Gold Standard in Neurodegenerative Disease – BioBuzz

Clene Nanomedicine, one of Marylands Future 2020 companies, closed a merger with Tottenham Acquisition I Limited that provided the company with a slot on the Nasdaq Exchange and $31.9 million that will be used to advance the companys gold nanocrystal-based treatment for neurodegenerative diseases.

Clene is developing a new class of drugs called bioenergetic nanotherapeutics that harnesses the properties of gold nanocrystals, which are used to amplify bioenergetic reactions in patients in order to drive intracellular biological reactions. Chief Executive Officer Rob Etherington said bioenergetic nanotherapeutics are a clean break from pharmaceutical drug development that uses classical synthetic chemistry.

Using gold as a therapeutic is not a new concept. Gold-salt injections were historically used to treat rheumatoid arthritis decades ago, but were dropped due to health concerns and toxicity issues.

Funds from the merger with Tottenham, a special purpose acquisition company (SPAC), will be used to advance Clenes lead asset, CNM-Au8, a bioenergetic nanocatalyst under development as an add-on treatment for neurodegenerative diseases like Parkinsons disease, multiple sclerosis and Amyotrophic Lateral Sclerosis (ALS). CNM-Au8 is designed to catalyze biocelluar reactions and so far the company has seen the asset live up to its promise in clinical studies.

Although Clenes corporate offices are in Utah, the bulk of the companys research is conducted in Cecil County, Maryland. Approximately three-fourths of the companys employees are located in Maryland, Etherington said. The company conducts all R&D and manufacturing in Maryland, as well as its commercialization activities.

Maryland has been a perfect anchor for Clene, Etherington said.

While the company raised a significant amount of funds through the merger, Etherington said Maryland was one of the companys earliest financial backers through a loan arrangement. He said predicted his company and the state will continue to have a strong relationship in the years ahead.

The state has been a true supporter. It was also gratifying to be recognized as a Future 2020 company because we have built a home in Maryland, he said.

Etherington predicted the next 18 months will be important for the company due to a planned readout of the companys asset. The funds raised in the merger will be used to carry CNM-Au8 into Phase II and Phase III studies that aim to address neurodegenerative diseases of high unmet medical need, such as multiple sclerosis, Parkinsons disease and amyotrophic lateral sclerosis (ALS). CNM-Au8 is currently being assessed in a Phase II study for the treatment of chronic optic neuropathy in patients with multiple sclerosis and is in Phase II and Phase III studies for disease progression in patients with ALS. Both Phase II studies are expected to read out this year. Its also being studied in a Phase II Parkinsons disease trial. The company reported interim data from the study last year and additional data is expected in the first half of 2021. Clene also plans to launch an additional Phase IIb Parkinsons disease efficacy study with CN<-Au8 by the end of the year.

While CNM-Au8 is showing promise, its not designed to replace any medication currently being used by a patient for one of these diseases. Rather, its designed to work alongside those medications. Clenes compound is designed to enhance the intracellular biological actions necessary to repair and reverse neuronal damage, Etherington said.

Clenes gold nanocrystals are grown in treated water and patients drink the product when it is ready. By combining the nanocrystal therapy with ongoing treatments for these neurodegenerative diseases, Etherington said the goal is to reverse neurodegeneration.

We want to let the cell take care of its own housekeeping and enhance whats naturally occurring in the central nervous system, he said.

In addition to the gold nanocrystal compound, Clene is developing a silver and zinc asset for COVID-19. CNM-AgZn17 is Clenes second key asset intended for broad anti-viral and anti-microbial use. A Phase II study is planned in Brazil to treat acutely symptomatic non-hospitalized patients with COVID-19.

Alex Keown is a freelance journalist who writes about a variety of subjects including the pharma, biotech, and life science industries. Prior to freelancing, Alex has served as a staff writer and editor for several publications.

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Clene Nanomedicine Aims to be the Gold Standard in Neurodegenerative Disease - BioBuzz

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Clene Nanomedicine Receives Patent Notice of Allowance in the United States for using Gold Nanocrystals for the Treatment of Multiple Sclerosis -…

SALT LAKE CITY, Jan. 19, 2021 (GLOBE NEWSWIRE) -- Clene Inc. (NASDAQ: CLNN) (along with its subsidiaries, Clene) today announced that its wholly-owned subsidiary Clene Nanomedicine, Inc., a clinical-stage biopharmaceutical company, was issued a Notice of Allowance from the U.S. Patent and Trademark Office (USPTO) for its invention for using its patented clean-surfaced gold nanocrystals for treating patients with multiple sclerosis (MS).

Clene notes that its previously patented gold nanocrystals have surfaces that are substantially free from organic impurities and are therefore clean relative to surfaces of gold nanoparticles made by other processes. The allowed application discloses that these gold nanocrystals can be suspended in water and can be taken orally, for example, by a person with MS.

As the leading developer of clean surfaced nanocrystal therapeutics for humans, we continue to expand our patent estate and are pleased to receive this latest Notice of Allowance from the USPTO. While the only approved treatments for MS today are immunomodulators, we see an opportunity to treat MS through a completely different mechanism of action utilizing the therapeutic bioenergetic effects of catalytic gold nanocrystals, such as CNM-Au8. This Notice of Allowance comes as we are conducting two Phase 2 studies of our lead drug candidate CNM-Au8 in the treatment of MS, stated Rob Etherington, President and CEO of Clene.

MS affects anestimated 1 million people in the U.S. and 2.5 million worldwide, with a treatment market valued at $23 billionglobally.

New data is expected from Clenes REPAIR-MS Phase 2 study of CNM-Au8 in the second half of 2021. Prior interim results from this study showed CNM-Au8 was associated with improvements across key central nervous system (CNS) bioenergetic metabolites.

Clenes VISIONARY-MS Phase 2 study is evaluating the efficacy and safety of CNM-Au8 as a remyelinating and neuro-reparative treatment in stable relapsing MS patients with chronic visual impairment. Interim data from the Phase 2 VISIONARY-MS trial demonstrated notable, exposure-related median improvements in the primary endpoint. Completion of enrollment is expected by the end of 2021.

Clenes worldwide patent portfolio in the new field of clean-surfaced nanocrystal therapeutics includes over 100 patents issued and allowed, with around 30 more pending. The issued patents cover state of matter claims for suspensions and solutions, as well as processes for making the materials, devices for conducting the unique electro-crystal chemistry processes, and methods of using the novel materials, such as in this instance of using clean-surfaced gold nanocrystals for treating patients with MS.

About CNM-Au8

CNM-Au8 is a concentrated, aqueous suspension of clean-surfaced faceted gold nanocrystals that act catalytically to support important intracellular biological reactions. CNM-Au8 consists solely of gold nanoparticles, composed of clean-surfaced, faceted, geometrical crystals held in suspension in sodium bicarbonate buffered, pharmaceutical grade water. CNM-Au8 has demonstrated safety in Phase 1 studies in healthy volunteers and has shown both remyelination and neuroprotective effects in multiple preclinical (animal) models. Preclinical data, both published in peer-reviewed journals and presented at scientific congresses, demonstrate that treatment of neuronal cultures with CNM-Au8 improves survival of neurons, protects neurite networks, decreases intracellular levels of reactive oxygen species and improves mitochondrial capacity in response to cellular stresses induced by multiple disease-relevant neurotoxins. Oral treatment with CNM-Au8 improved functional behaviors in rodent models of ALS, MS and Parkinsons disease versus vehicle (placebo). CNM-Au8 is currently being tested in a Phase 2 clinical study for the treatment of chronic optic neuropathy in patients with MS, in addition to Phase 2 and Phase 3 clinical studies for disease progression in patients with ALS.

About Clene

Clene is a clinical-stage biopharmaceutical company focused on the development of unique therapeutics for neurodegenerative diseases. Clene has innovated a novel nanotechnology drug platform for the development of a new class of orally administered neurotherapeutic drugs. Clene has also advanced into the clinic an aqueous solution of ionic zinc and silver for anti-viral and anti-microbial uses. Founded in 2013, the company is based in Salt Lake City, Utah with R&D and manufacturing operations located in North East, Maryland. For more information, please visit http://www.clene.com.

Forward-Looking Statements

This press release contains "forward-looking statements" within the meaning of the "safe harbor" provisions of the Private Securities Litigation Reform Act of 1995. Clene's actual results may differ from its expectations, estimates and projections and consequently, you should not rely on these forward-looking statements as predictions of future events. Words such as "expect," "estimate," "project," "budget," "forecast," "anticipate," "intend," "plan," "may," "will," "could," "should," "believes," "predicts," "potential," "might" and "continues," and similar expressions are intended to identify such forward-looking statements. These forward-looking statements involve significant known and unknown risks and uncertainties, many of which are beyond Clenes control and could cause actual results to differ materially and adversely from expected results. Factors that may cause such differences include Clenes ability to demonstrate the efficacy and safety of its drug candidates; the clinical results for its drug candidates, which may not support further development or marketing approval; actions of regulatory agencies, which may affect the initiation, timing and progress of clinical trials and marketing approval; Clenes ability to achieve commercial success for its marketed products and drug candidates, if approved; Clenes ability to obtain and maintain protection of intellectual property for its technology and drugs; Clenes reliance on third parties to conduct drug development, manufacturing and other services; Clenes limited operating history and its ability to obtain additional funding for operations and to complete the licensing or development and commercialization of its drug candidates; the impact of the COVID-19 pandemic on Clenes clinical development, commercial and other operations, as well as those risks more fully discussed in the section entitled Risk Factors in Clenes recently filed registration statement on Form S-4, as well as discussions of potential risks, uncertainties, and other important factors in Clenes subsequent filings with the U.S. Securities and Exchange Commission. Clene undertakes no obligation to release publicly any updates or revisions to any forward-looking statements to reflect any change in its expectations or any change in events, conditions or circumstances on which any such statement is based, subject to applicable law. All information in this press release is as of the date of this press release. The information contained in any website referenced herein is not, and shall not be deemed to be, part of or incorporated into this press release.

Media ContactAndrew MielachLifeSci Communications(646) 876-5868amielach@lifescicomms.com

Investor ContactBruce MackleLifeSci Advisors, LLC(929) 469-3859bmackle@lifesciadvisors.com

Source: Clene Inc.

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Clene Nanomedicine Receives Patent Notice of Allowance in the United States for using Gold Nanocrystals for the Treatment of Multiple Sclerosis -...

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Recent Advances in Nanomedicine for the Diagnosis and Treatment of Prostate Cancer Bone Metastasis – DocWire News

This article was originally published here

Molecules. 2021 Jan 13;26(2):E384. doi: 10.3390/molecules26020384.

ABSTRACT

Patients with advanced prostate cancer can develop painful and debilitating bone metastases. Currently available interventions for prostate cancer bone metastases, including chemotherapy, bisphosphonates, and radiopharmaceuticals, are only palliative. They can relieve pain, reduce complications (e.g., bone fractures), and improve quality of life, but they do not significantly improve survival times. Therefore, additional strategies to enhance the diagnosis and treatment of prostate cancer bone metastases are needed. Nanotechnology is a versatile platform that has been used to increase the specificity and therapeutic efficacy of various treatments for prostate cancer bone metastases. In this review, we summarize preclinical research that utilizes nanotechnology to develop novel diagnostic imaging tools, translational models, and therapies to combat prostate cancer bone metastases.

PMID:33450939 | DOI:10.3390/molecules26020384

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NanoView Biosciences Strengthens Executive Team – BioSpace

BOSTON, Jan. 13, 2021 /PRNewswire/ -- NanoView Biosciences, Inc., the leader in single exosome characterization, today announced the expansion of its management team with the appointment of David Hanlon, Ph.D. to the position of Vice President, Strategic Collaborations. The addition is a key part of the company's strategy to create and expand strategic partnerships within the life sciences, pharmaceutical and diagnostics markets.

Dr. Hanlon brings over 20 years of product development and commercialization experience to NanoView, most recently serving as Vice President of Strategic Collaborations at Quanterix. Over the last 12 years he has led the strategic marketing efforts and established an assay service laboratory to support new applications and drive commercial interest. He has held leadership roles in several life sciences and diagnostics companies, including Hologic (formerly Cytyc), where he directed multiple biomarker programs to develop diagnostic tests for cervical cancer. Dr. Hanlon received his Ph.D. in Biochemistry from the University of Illinois, Urbana-Champaign and B.S. from Worcester Polytechnic Institute.

"We are very excited to welcome David Hanlon to NanoView as we enter a significant growth phase for the Company," said Jerry Williamson, CEO of NanoView Biosciences. "His expertise in establishing and expanding strategic relationships for life science technology providers will accelerate the success of NanoView and we look forward to his contribution on our executive team."

Commenting on his new role, Dr. Hanlon added, "I am very excited to be joining NanoView, a ground-breaking organization that has developed a novel platform solution to characterize extracellular vesicles in human disease. I look forward to working with the leadership team as the organization continues to expand its product and service offerings to help researchers and clinicians leverage the potential of exosomes in this era of personalized medicine."

NanoView has created ExoView, a unique and proprietaryplatform which provides high-resolution sizing, counting and phenotyping of exosomes and viral vectors at the single vesicle level. Understanding the molecules carried by extracellular vesicles has potential for diagnostic, prognostic, and therapeutic use for a broad range of diseases. ExoView requires low sample input, no extensive sample preparation or purification, and minimal hands-on time. It can be used directly with complex biological samples.

The ExoView platform is designed For Research Use Only. It is not for use in diagnostic procedures.

About NanoView Biosciences

NanoView Biosciences, a Boston-based, privately-held company, is focused on enabling worldwide life science researchers to better understand the biological role of extracellular vesicles, including exosomes and viral vectors, and their potential use as biomarkers for improving the diagnosis, prognosis, treatment, and monitoring of disease. The Company's proprietary product, the ExoView platform, was designed to fully characterize exosomes and other extracellular vesicles for use in research and in the implementation of precision nanomedicine. ExoView is a high-throughput, cost-effective analysis platform that is easy to use and does not require purification or large sample volumes to accurately analyze exosomes.

http://www.nanoviewbio.com

View original content:http://www.prnewswire.com/news-releases/nanoview-biosciences-strengthens-executive-team-301207135.html

SOURCE NanoView Biosciences

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Amyotrophic Lateral Sclerosis Market Anticipates Sizeable Growth at a CAGR of 28% during the forecast period (2020-2030) with the US expected to top…

LAS VEGAS, Jan. 18, 2021 /PRNewswire/ -- The surge in the Amyotrophic Lateral Sclerosis (ALS) market can be attributed to a vast pipeline, initiation of HEALEY ALS platform trials, mutation-specific treatment development in the pipeline, a need for a curative treatment option, increase in the ALS prevalence, effective drug approvals in the past 3-4 years, improvisations in market regulatory guidelines, growing awareness about the disease, along with geographical expansions and meaningful collaborations in the ALS pharma industry.

Amyotrophic Lateral Sclerosis Marketreport offers detailed coverage of the disease, its available treatment options, patient pool, and diagnostic modalities. The report lays down the comprehensive insights into market outlook, upcoming pipeline therapies, and unmet needs along with the major collaboration and funding opportunities in the ALS market landscape.

Some of the highlights of the Amyotrophic Lateral Sclerosis marketreport:

Know more about the report highlights @Amyotrophic Lateral Sclerosis Market Landscape and Forecast

Amyotrophic Lateral Sclerosis, commonly known as Lou Gehrig's disease, is a group of progressive, rare neurological diseases, which are a result of gradual deterioration and death of motor neurons. Over time the muscles weaken, start to twitch (called fasciculations), and waste away (atrophy), eventually, leading the brain to lose its ability to initiate and control voluntary movement.

ALS can be either sporadic or genetic. The sporadic type is the most common and can affect anyone. The genetic or familial type is rarer. On the basis of mutation, ALS patients can exhibit mutations such as C9ORF72, SOD1, along with others including TARDBP, FUS, OPTN, ANG, etc., and non-mutated/unidentified mutations. The diagnosed Amyotrophic Lateral Sclerosis prevalent cases in the 7MM are estimated to be 48,112 in 2020, which is further expected to increase by 2030.

The Amyotrophic lateral sclerosis epidemiology section of the report proffers insights into the historical and current Amyotrophic lateral sclerosis (ALS) patient pool and forecasted trends for 7MM. ALS report provides historical as well as forecasted epidemiology segmentation for the study period 2017-30:

Visit, Amyotrophic Lateral Sclerosis Epidemiology, for more information

Amyotrophic Lateral Sclerosis Therapeutic Market Landscape

The Amyotrophic Lateral Sclerosis treatment landscape includes multidisciplinary care, such as physical therapy, speech therapy, dietary counselling, heat or whirlpool therapy and others. Approved drugs in the ALS market comprises Riluzole, Nuedexta, Radicava, and Tiglutik. Rilutek (Riluzole; Covis Pharmaceuticals) is indicated for the treatment of ALS and the mechanism by which riluzole exerts its therapeutic effects in patients with ALS is still unknown. Another approved drug Tiglutik (riluzole), an oral suspension, is the first and only easy-to-swallow thickened riluzole liquid for ALS and is administered twice daily via an oral syringe. On the other hand, Exservan is an oral Riluzole film used to treat ALS. Nuedexta is available as a combination drug containing dextromethorphan hydrobromide and quinidine sulfate indicated for the treatment of pseudobulbar affect. Radicava (Edaravone; Mitsubishi Tanabe Pharma) is another treatment option for the ALS. During clinical trials, it has shown to decrease the decline of the physical functions, however, it is capable of causing adverse effects owing to its constituent sodium bisulfite.

Request for sample @Amyotrophic Lateral Sclerosis Treatment Marketfor rich insights into treatment modalities

Medications are also be given to manage symptoms of ALS, including pain, muscle cramps, stiffness, excess saliva and phlegm, and the pseudobulbar affect (involuntary or uncontrollable episodes of crying and/or laughing, or other emotional displays). Drugs also are available to help individuals with pain, depression, sleep disturbances, and constipation. There also exists a variety of tools and mechanical devices such as non-invasive ventilation (NIV) and mechanical ventilation for breathing support to help with ALS such as splints, reach extenders, and grab-bars.

However, the Amyotrophic Lateral Sclerosis therapeutic market lacks a curative approach that can halt or reverse the progression of the disease. Even though Radicava can effectively delay physical disability in ALS patients, however, its high pricing may pose a threat to the overall patient compliance of the drug. Furthermore, the lack of epidemiological data of the disease in the pediatric population hinders the development of novel therapies focused on children below 18 years of age.A poor understanding of the mutations, which form a major cause of the ALS, dims down the ongoing speed of the development of the pipeline therapies. Thus, there is a need for better research opportunities in the domain to facilitate a better understanding of the disease and development of potential novel therapies.

Amyotrophic Lateral Sclerosis Pipeline Therapies

Know more about the top emerging therapies in the ALS market @ Amyotrophic Lateral Sclerosis Drug Pipeline

Amyotrophic Lateral Sclerosis Market Forecast

DelveInsight estimates that approval of novel Amyotrophic Lateral Sclerosis pipeline therapies is expected to give the ALS market size a much-needed boost in the foreseeable future. Although, most of the therapies are going to get commercialized in combination with Riluzole and Radicava. In addition to this, other emerging therapies are under trials while the patient is already following their treatment regimen of Riluzole. Thus, it can be said that the pipeline therapies will be sharing the market share with the available market therapies.

Majority of emerging drugs are focusing on delaying the disease progression, and are not curative. Thus, the ALS market presents immense opportunities for pharmaceutical companies to explore and exploit their ground-breaking therapies and occupy the maximum potential. Here, gene and enzyme replacement therapies, which promise to cure the disease completely, automatically get the upper hand here. Several pharmaceutical companies are investigating novel approaches and joining hands with others to expedite the development of ALS treatment options. Recently, Catalent and BrainStorm entered into an agreement to manufacture NurOwn. Cytokinetics and Astellas entered into the Fast Skeletal Regulatory Activator Agreement, allowing Cytokinetics to exercise exclusive control over the future development and commercialization of reldesemtiv and other FSRA compounds and products. Besides agreements, acquisition spree in the ALS market is quite dominant as well. UCB acquired Ra Pharmaceuticals and got zilucoplan, which is currently in phase III, to its vast portfolio. Similarly, Novus Therapeutics completed the acquisition of Anelixis Therapeutics, which is conducting a phase II clinical trial of AT-1501 for ALS.

Although several clinical trials are ongoing, however, an unsettling thing that might be a snag in the steady growth of the Amyotrophic Lateral Sclerosis market is the quite unpropitious success rate of the clinical trials. The management of ALS is extremely expensive. Expensive drugs like Radicava pose a burden for ALS patients as not everyone can get access to the drug due to its high cost. Thus, the cost-effectiveness of the upcoming therapies is also expected to play a major part in their approval.

Know more about the collaboration and funding ongoing @ Major collaborations in Amyotrophic Lateral Sclerosis Market

Scope of the report

Geography Coverage: The United States, EU5 (Germany, France, Italy, Spain, and the United

Kingdom), Japan.Study Period: 3-year historical and 11-year forecasted analysis (2017-2030).Amyotrophic Lateral Sclerosis Market Segmentation: By Geographies, By Therapies.Key Players Involved: Orphazyme, Biogen/Ionis Pharmaceuticals, MediciNova, AB Sciences, Novus Therapeutics, Seelos Therapeutics, Brainstorm-Cell Therapeutics, Amylyx Pharmaceuticals, Gilead Sciences, PTC Therapeutics, GlaxoSmithKline, Clene Nanomedicine, Alexion Pharmaceuticals, Biohaven Pharmaceuticals, UCB Pharma, and several others.Analysis: Comparative and conjoint analysis of Amyotrophic Lateral Sclerosis Emerging therapies, Attribute AnalysisAmyotrophic Lateral Sclerosis Market trends, pipeline analysis across different stages of development (Phase III and Phase II), and Amyotrophic Lateral Sclerosis Market size by therapies.Tools used: SWOT analysis, Porter's Five Forces, PESTLE analysis, BCG Matrix analysis methods.Case StudiesKOL's ViewsAnalyst's Views

Table of Contents

1

Key Insights

2

Executive Summary of Amyotrophic lateral sclerosis

3

Competitive Intelligence Analysis for Amyotrophic Lateral Sclerosis

4

ALS Market Overview at a Glance

5

ALS - Disease Background and Overview

6

Amyotrophic Lateral Sclerosis Patient Journey

7

Amyotrophic Lateral Sclerosis Epidemiology and Patient Population

8

ALS Treatment Algorithm, Current Treatment, and Medical Practices

9

Amyotrophic Lateral Sclerosis Unmet Needs

10

Key Endpoints of Amyotrophic lateral sclerosis Treatment

11

ALS Marketed Products

12

Amyotrophic Lateral Sclerosis Emerging Therapies

13

ALS Seven Major Market Analysis

14

Attribute analysis

15

Amyotrophic Lateral Sclerosis (7MM) Market Outlook

16

Access and Reimbursement Overview of ALS

17

KOL Views

18

Amyotrophic Lateral Sclerosis Market Drivers

19

ALS Market Barriers

20

Appendix

21

DelveInsight Capabilities

22

Disclaimer

23

About DelveInsight

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